Quinazoline derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly discloses a quinazoline derivative as well as a preparation method and application thereof. The structure of the quinazoline derivative is shown as a formula I. The invention provides a novel quinazoline derivative capable of efficiently inhibiting LSD1 activity, the preparation process is simple and easy to implement, a good effect of resisting colorectal cancer cell proliferation is shown on 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 biomedical technology, specifically relating to a quinazoline derivative, its preparation method, and its application. Background Technology

[0002] Lysine-specific demethylase 1 (LSD1, also known as KDM1A) is a key molecule in the field of epigenetic regulation. It participates in the precise regulation of chromatin remodeling and gene transcription through the dynamic control of histone H3K4 and H3K9 methylation modifications. As the first histone demethylase identified, LSD1 dysfunction is closely related to multi-stage pathological processes in malignant tumors, making it an important potential target for current anti-tumor drug development.

[0003] In the initial stages of tumorigenesis, abnormally high expression of LSD1 can induce cellular carcinogenesis through epigenetic modification imbalance. Studies have shown that in gastric cancer tissues, LSD1 interacts with the transcriptional repressor complex to specifically demethylate H3K4me2 in the promoter region of the tumor suppressor gene p53, leading to transcriptional silencing and thus releasing the cell cycle checkpoint constraint. In pancreatic cancer, LSD1-mediated H3K9me2 demethylation can activate the expression of key genes in the Hedgehog signaling pathway (such as Gli1), driving the malignant transformation of pancreatic ductal epithelial cells. These findings reveal the epigenetic driving role of LSD1 in tumorigenesis.

[0004] LSD1 plays an irreplaceable role in maintaining the tumor proliferation phenotype. In an ovarian cancer model, LSD1 accelerates the cell cycle by activating the transcription of genes encoding cyclin-dependent kinases (CDK2 / 4), enabling tumor cells to acquire sustained proliferative capacity. In glioblastoma, LSD1 forms a complex with stem cell transcription factors such as SOX2, maintaining the self-renewal characteristics of tumor stem cells, which is closely related to rapid tumor growth and treatment resistance. Clinical sample analysis shows that tumor patients with high LSD1 expression often have shorter progression-free survival, further confirming its clinical significance as a driver of tumor proliferation.

[0005] Tumor invasion and metastasis are major causes of treatment failure, and LSD1 plays a crucial regulatory role in this process. In breast cancer, LSD1 demethylates H3K4me1 / 2, relieving the inhibition of key epithelial-mesenchymal transition (EMT) transcription factors such as Snail and Slug, thus promoting tumor cells' acquisition of mesenchymal phenotype and migration ability. In liver cancer, LSD1 can enhance the tumor cells' ability to degrade the extracellular matrix by regulating the expression of matrix metalloproteinase family members (such as MMP-14), creating conditions for distant metastasis. These studies suggest that targeting LSD1 may be an effective strategy for inhibiting tumor metastasis.

[0006] Furthermore, aberrant activation of LSD1 is closely related to the development of tumor treatment resistance. In non-small cell lung cancer, LSD1 increases the efflux of chemotherapeutic drugs by epigenetically regulating the expression of ABC transporter family genes (such as ABCG2). In endocrine therapy for breast cancer, LSD1-mediated demethylation of the estrogen receptor (ER) promoter can lead to persistent ER activation, causing tumor cells to develop resistance to endocrine therapy drugs such as tamoxifen. Simultaneously, LSD1 can also affect the immunosuppressive state of the tumor microenvironment by regulating the expression of immune checkpoint molecules (such as PD-L1), thereby reducing the efficacy of immunotherapy.

[0007] Given the crucial regulatory role of LSD1 in multiple key stages of tumorigenesis, proliferation, metastasis, and drug resistance, the development of specific inhibitors targeting LSD1 has become an important direction in anti-tumor drug development. LSD1-targeted therapy strategies hold promise for providing a new breakthrough in the precision treatment of malignant tumors and have significant clinical translational value. Summary of the Invention

[0008] The purpose of this application is to solve at least one technical problem in the prior art. Based on this, the first aspect of this application provides a quinazoline derivative with the structure shown in Formula I.

[0009]

[0010] Among them, R 1 Selected from hydrogen, cyclopropyl, methyl, phenyl, N-methylpiperazine, 4-methylhexahydropyridine, thiazole; R 2 Selected from hydrogen, chlorine, bromine, fluorine, and methoxy.

[0011] As a further preferred embodiment, the structure of the above-mentioned quinazoline derivative is any one of 4a-4n; more preferably, the structure of the above-mentioned quinazoline derivative is 4b.

[0012]

[0013] A second aspect of this application also provides a method for preparing the above-mentioned quinazoline derivative, comprising the following synthetic route:

[0014]

[0015] 2. Sulfonamide compound was obtained by reacting 1H-indole-5-sulfonyl chloride 1 (commercially available) with a substituted amino group under triethylamine conditions. The final product was then prepared by reacting it with a 2-chloroquinazoline 3 derivative. The synthetic method described in this application is simple, uses readily available starting materials, is easy to prepare, and exhibits good functional group compatibility.

[0016] A third aspect of this application also provides an LSD1 inhibitor, comprising the above-described quinazoline derivative or a pharmaceutically acceptable salt thereof.

[0017] A fourth aspect of this application also provides a pharmaceutical composition comprising the above-mentioned quinazoline derivative or a pharmaceutically acceptable salt thereof. The pharmaceutical composition includes an excipient; the excipient is at least one selected from gum arabic, syrup, lanolin, and starch. This excipient is stable, has no incompatibility with the active pharmaceutical ingredient, does not produce side effects, does not affect efficacy, is not easily deformed, cracked, moldy, or infested by insects at room temperature, is harmless to the human body, has no physiological effect, does not react chemically or physically with the active pharmaceutical ingredient, and does not affect the content determination of the active pharmaceutical ingredient.

[0018] The fifth aspect of this application also provides the use of the above-mentioned quinazoline derivative in the preparation of reagents for diagnosing and / or treating diseases characterized by LSD1 overexpression. The preferred disease is colorectal cancer.

[0019] The beneficial effects of this application are as follows: This application provides a novel quinazoline derivative with highly efficient LSD1 inhibition activity. The preparation process is simple and easy. 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

[0020] The following will provide a clear and complete description of the concept and technical effects of this application in conjunction with embodiments, so as to fully understand the purpose, solution and effects of this application. It should be noted that, unless otherwise specified, the embodiments and features in the embodiments of this application can be combined with each other.

[0021] The synthetic routes for the quinazoline derivatives 4a-4n involved in the following content are as follows:

[0022]

[0023] The structures of quinazoline derivatives 4a-4n are shown below:

[0024]

[0025] The structure of the corresponding compound 2a-2i is shown below:

[0026]

[0027] The preparation process of compound 2 is as follows: 1H-indole-5-sulfonyl chloride 1 (215.0 mg, 1.0 mmol, 1.0 eq) was dissolved in dichloromethane. Under nitrogen protection, the corresponding amines (1.2 mmol, 1.2 eq) and triethylamine (202.0 mg, 2.0 mmol, 2.0 eq) were added sequentially. After the reaction of the raw materials was completed, saturated brine was added to quench the reaction. The mixture was extracted with dichloromethane, and the organic phases were combined, dried and concentrated with anhydrous sodium sulfate, and purified by column chromatography to obtain compounds 2a-2i.

[0028] In the preparation of compound 2a (212.4 mg, 90% yield), the amine added was cyclopropylamine (68.4 mg, 1.2 mmol, 1.2 eq), and the characterization results were as follows: 1 H NMR(400MHz,Chloroform-d)δ8.28–8.21(m,2H),7.70(dd,J=7.5,1.5Hz,1H),7.56(d,J=7.5Hz,1H),7.18(d,J=7 .5Hz,1H),6.66(dd,J=7.5,1.4Hz,1H),4.06(s,1H),2.25(p,J=7.0Hz,1H),0.64–0.45(m,2H),0.33–0.13(m,2H). 13 CNMR(100MHz,Chloroform-d)δ139.19,135.61,125.31,124.20,124.00,121.88,111.82,102.16,24.46,8.77.

[0029] The amine added during the preparation of compound 2b (256.7 mg, 92% yield) was N-methylpiperazine (120.1 mg, 1.2 mmol, 1.2 eq), and the characterization results were as follows: 1 H NMR(400MHz,Chloroform-d)δ8.28–8.20(m,2H),7.68(dd,J=7.5,1.5Hz,1H),7.56(d,J=7.5Hz,1H),7.18 (d,J=7.5Hz,1H),6.66(dd,J=7.4,1.5Hz,1H),3.10(t,J=5.1Hz,4H),2.50(t,J=5.1Hz,4H),2.27(s,3H). 13C NMR (100MHz, Chloroform-d) δ139.10,134.27,125.31,124.91,124.28,123.31,113.26,102.16,51.60,45.47,45.04.

[0030] The amine added during the preparation of compound 2c (185.9 mg, yield 83%) was dimethylamine (54.1 mg, 1.2 mmol, 1.2 eq), and the characterization results were as follows: 1 H NMR(400MHz,Chloroform-d)δ8.27–8.19(m,2H),7.69(dd,J=7.5,1.5Hz,1H),7.56 (d,J=7.5Hz,1H),7.18(d,J=7.5Hz,1H),6.66(dd,J=7.4,1.5Hz,1H),2.72(s,6H). 13 C NMR (100MHz, Chloroform-d) δ138.97,134.27,125.31,125.00,124.43,123.99,113.75,102.16,37.99.

[0031] The amine added during the preparation of compound 2d (165.9 mg, yield 79%) was a 40% aqueous solution of methylamine (93.3 mg, 1.2 mmol, 1.2 eq). The characterization results are as follows: 1 H NMR (400MHz, Chloroform-d) δ8.29–8.20 (m, 2H), 7.72 (dd, J=7.4, 1.5Hz, 1H), 7.56 (d, J= 7.5Hz,1H),7.18(d,J=7.5Hz,1H),6.66(dd,J=7.5,1.6Hz,1H),4.50(s,1H),2.55(s,3H). 13 C NMR (100MHz, Chloroform-d) δ140.78,135.78,125.31,124.77,123.85,122.65,112.15,102.16,28.96.

[0032] The amine added during the preparation of compound 2e (231.2 mg, yield 85%) was aniline (111.7 mg, 1.2 mmol, 1.2 eq), and the characterization results are as follows: 1H NMR(400MHz,Chloroform-d)δ8.39(d,J=1.4Hz,1H),8.23(s,1H),7.65(dd,J=7.5,1.5Hz,1H),7.50(d,J=7.5Hz,1H),7.34–7. 26(m,2H),7.18(d,J=7.5Hz,1H),7.12(t,J=7.5Hz,2H),6.80(tt,J=7.5,2.0Hz,1H),6.70(dd,J=7.5,1.5Hz,1H),6.09(s,1H). 13 C NMR (100MHz, Chloroform-d) δ138.35,135.33,135.20,129.15,125.31,124.26,124.05,123.53,121.38,119.69,111.61,102.16.

[0033] The amine added during the preparation of compound 2f (200.2 mg, yield 72%) was 4-methylhexahydropyridine (118.9 mg, 1.2 mmol, 1.2 eq), and the characterization results were as follows: 1 H NMR(400MHz,Chloroform-d)δ8.36–8.13(m,2H),7.73(dd,J=7.4,1.6Hz,1H),7.56(d,J=7.5Hz,1H),7.18(d,J=7.4Hz,1H),6.66(dd,J=7 .4,1.5Hz,1H),4.02–3.82(m,2H),3.11–2.90(m,2H),1.70(dddd,J=13.0,8.5,7.2,4.5Hz,2H),1.47–1.25(m,3H),0.91(d,J=6.2Hz,3H). 13 C NMR (100MHz, Chloroform-d) δ139.10,134.27,125.31,124.91,124.28,123.31,113.26,102.16,45.16,33.97,30.17,22.01.

[0034] The amine added during the preparation of compound 2g (203.7mg, yield 73%) was 2-aminothiazole (120.2mg, 1.2mmol, 1.2eq), and the characterization results were as follows: 1H NMR(400MHz,Chloroform-d)δ9.00(s,1H),8.23(s,2H),7.76(dd,J=7.5,1.5Hz,1H),7.60(d,J=7.5Hz ,1H),7.25(d,J=7.5Hz,1H),7.18(d,J=7.5Hz,1H),6.83(d,J=7.5Hz,1H),6.59(dd,J=7.4,1.5Hz,1H). 13 C NMR (100MHz, Chloroform-d) δ162.48,138.35,135.33,125.55,125.31,124.05,123.53,121.38,111.61,108.62,102.16.

[0035] The amine added during the preparation of compound 2h (225.2 mg, yield 81%) was cycloheximine (120.2 mg, 1.2 mmol, 1.2 eq), and the characterization results were as follows: 1 H NMR(400MHz,Chloroform-d)δ8.29–8.18(m,2H),7.72(dd,J=7.5,1.5Hz,1H),7.55(d,J=7.5Hz,1H),7.18(d,J =7.5Hz,1H),6.66(dd,J=7.4,1.5Hz,1H),3.21–3.09(m,4H),1.60(q,J=2.7,2.1Hz,4H),1.48(p,J=5.5Hz,4H). 13 C NMR (100MHz, Chloroform-d) δ139.10,134.27,125.31,124.91,124.28,123.31,113.26,102.16,48.44,28.16,26.66.

[0036] The amine added during the preparation of compound 2i (300.5 mg, yield 81%) was N-(4-aminophenyl)-2,2-dimethylpropionamide (230.7 mg, 1.2 mmol, 1.2 eq), and the characterization results were as follows: 1H NMR(400MHz,Chloroform-d)δ9.50(s,1H),8.40(t,J=1.6Hz,1H),8.23(s,1H),7.86(dd,J=7.5,1.5Hz,1H),7.66( d,J=7.5Hz,1H),7.49–7.43(m,2H),7.18(d,J=7.5Hz,1H),6.71(dd,J=8.1,1.9Hz,3H),6.20(s,1H),1.40(s,9H). 13 C NMR(100MHz,Chloroform-d)δ177.25,138.35,135.33,135.14,133.64,125. 31,125.10,124.18,124.05,123.53,121.38,111.61,102.16,39.54,27.60.

[0037] Example 1

[0038] A quinazoline derivative, the structure of which is shown in 4a; its preparation method includes the following steps:

[0039] 1H-indole-5-sulfonamide (196.2 mmol, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 2,7-dichloroquinazoline (199.0 mg, 1.0 mmol, 1.0 eq) was added. 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 4a (186.2 mg, yield 52%). 1 H NMR(400MHz,Chloroform-d)δ9.56(d,J=1.4Hz,1H),8.41(t,J=1.5Hz,1H),8.21(d,J=7.5Hz,1H),8.14(d,J=1.4Hz,1H),7.9 0(ddd,J=13.2,7.5,1.5Hz,2H),7.81(d,J=7.5Hz,1H),7.69(dd,J=7.5,1.5Hz,1H),6.83(dd,J=7.5,1.6Hz,1H),4.48(s,2H). 13C NMR (100MHz, Chloroform-d) δ155.1,153.6,150.3,138.9,136.4,135.4,129.3,128.2,127.0,126.4,125.6,125.2,121.6,120.8,112.0,109.6.

[0040] Example 2

[0041] A quinazoline derivative, the structure of which is shown in 4b; its preparation method includes the following steps:

[0042] Compound 2a (236.0 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 2,7-dichloroquinazoline (199.0 mg, 1.0 mmol, 1.0 eq) was added. 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 4b (163.2 mg, yield 41%). 1 HNMR(400MHz,Chloroform-d)δ9.40(d,J=1.7Hz,1H),8.25(t,J=2.1Hz,1H),8.17(dd,J=9.2,2.0Hz,1H),8.01(dd,J=8.7,2.3Hz,1H),7.87–7.81(m,2H) ,7.59(d,J=8.7Hz,1H),7.50(dd,J=9.1,2.2Hz,1H),7.11(d,J=9.1Hz,1H), 7.06(dd,J=6.8,2.2Hz,1H),2.61(dp,J=9.2,5.7Hz,1H),0.71–0.55(m,4H). 13 C NMR(100MHz,Chloroform-d)δ155.1,153.6,150.3,137.1,136.0,135.4,129. 3,128.2,127.0,126.7,126.5,125.2,122.7,121.6,112.0,109.5,26.3,8.7.

[0043] Example 3

[0044] A quinazoline derivative, the structure of which is shown in 4c; its preparation method includes the following steps:

[0045] Compound 2b (279.1 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0°C. After stirring for half an hour, 2,7-dichloroquinazoline (199.0 mg, 1.0 mmol, 1.0 eq) was added. 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 4c (163.2 mg, yield 37%). 1 HNMR(400MHz,Chloroform-d)δ9.40(d,J=1.8Hz,1H),8.24(t,J=2.2Hz,1H),8.17(dd,J=9.2,2.0Hz,1H),7.95(dd,J=8.9,2.3Hz,1H),7.87–7.81(m,2 H),7.64(d,J=9.1Hz,1H),7.50(dd,J=9.1,2.3Hz,1H),7.06(dd,J=6.7,2.2 Hz,1H),3.10(ddd,J=6.4,5.2,1.4Hz,4H),2.57–2.46(m,4H),2.27(s,3H). 13 C NMR(100MHz,Chloroform-d)δ155.1,153.6,150.3,137.6,135.4,134.7,129.3,1 28.2,127.0,126.8,126.4,125.2,123.3,121.6,112.0,110.3,52.0,45.5,45.4.

[0046] Example 4

[0047] A quinazoline derivative, the structure of which is shown in 4d; its preparation method includes the following steps:

[0048] Compound 2c (224.1 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 2,7-dichloroquinazoline (199.0 mg, 1.0 mmol, 1.0 eq) was added. 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 4d (142.8 mg, yield 37%). 1HNMR(400MHz,Chloroform-d)δ9.40(d,J=1.7Hz,1H),8.21(t,J=2.3Hz,1H),8.17(dd,J=9.2,2.1Hz,1H),7.90(dd,J=8.8,2 .2Hz,1H),7.87–7.81(m,2H),7.64(d,J=8.7Hz,1H),7.50(dd,J=9.1,2.3Hz,1H),7.06(dd,J=6.7,2.2Hz,1H),2.75(s,6H). 13 C NMR (100MHz, Chloroform-d) δ155.1,153.6,150.3,137.6,135.4,134.4,129.3,128.2,127.0,126.9,126.1,125.2,123.5,121.6,112.0,110.3,38.5.

[0049] Example 5

[0050] A quinazoline derivative, the structure of which is shown in 4e; its preparation method includes the following steps:

[0051] Compound 2d (210.0 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 2,7-dichloroquinazoline (199.0 mg, 1.0 mmol, 1.0 eq) was added. 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 4e (145.1 mg, yield 39%). 1 HNMR(400MHz,Chloroform-d)δ9.40(d,J=1.7Hz,1H),8.30(t,J=2.3Hz,1H),8.17(dd,J=9.2,2.1Hz,1H),7.99(dd,J=8.7,2.3Hz,1H),7.87 –7.81(m,2H),7.59(d,J=8.8Hz,1H),7.50(dd,J=9.1,2.3Hz,1H),7.06(dd,J=6.7,2.2Hz,1H),6.84(q,J=6.8Hz,1H),2.58(d,J=6.6Hz,3H). 13C NMR (100MHz, Chloroform-d) δ155.1,153.6,150.3,137.6,135.5,135.4,129.3,128.2,127.6,127.0,126.7,125.2,123.1,121.6,112.0,109.5,29.4.

[0052] Example 6

[0053] A quinazoline derivative, the structure of which is shown in 4f; its preparation method includes the following steps:

[0054] Compound 2e (272.1 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 2,7-dichloroquinazoline (199.0 mg, 1.0 mmol, 1.0 eq) was added. 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 4f (182.3 mg, yield 42%). 1 HNMR(400MHz,Chloroform-d)δ9.72(s,1H),9.40(d,J=1.8Hz,1H),8.40(t,J=2.2Hz,1H),8.17(dd,J=9.2,2.0Hz,1H),8.04(dd,J=8.7,2.3Hz,1H),7.87–7.8 1(m,2H),7.64(d,J=8.6Hz,1H),7.50(dd,J=9.1,2.2Hz,1H),7.32–7.25(m,2H ),7.23–7.17(m,2H),7.06(dd,J=6.7,2.2Hz,1H),6.92(tt,J=6.8,1.2Hz,1H). 13 C NMR(100MHz,Chloroform-d)δ155.1,153.6,150.3,137.5,136.8,135.5,135.4,129.3 ,129.1,128.2,127.0,126.7,126.2,125.2,125.2,122.7,121.6,120.9,112.0,109.7.

[0055] Example 7

[0056] A quinazoline derivative, the structure of which is shown in 4g; its preparation method includes the following steps:

[0057] Compound 2f (278.1 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0°C. After stirring for half an hour, 2,7-dichloroquinazoline (199.0 mg, 1.0 mmol, 1.0 eq) was added. 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 4 g of the compound (220.0 mg, 50% yield). 1 HNMR(400MHz,Chloroform-d)δ9.40(d,J=1.8Hz,1H),8.24(t,J=2.2Hz,1H),8.17(dd,J=9.2,2.0Hz,1H),7.95(d d,J=8.9,2.3Hz,1H),7.87–7.81(m,2H),7.64(d,J=9.1Hz,1H),7.50(dd,J=9.1,2.2Hz,1H),7.06(dd,J=6.7,2.2H z,1H),3.53(ddd,J=12.5,9.3,6.6Hz,2H),3.34(ddd,J=12.5,9.3,6.6Hz,2H),1.68(dddd,J=11.9,9.3,6.6,5.2H z,2H),1.52(ddtd,J=11.6,6.4,5.2,1.2Hz,1H),1.32(dddd,J=12.2,9.3,6.6,5.2Hz,2H),1.01(d,J=6.5Hz,3H). 13 C NMR(100MHz,Chloroform-d)δ155.1,153.6,150.3,137.6,135.4,134.5,129.3,128 .2,127.0,126.8,126.4,125.2,123.3,121.6,112.0,110.3,44.9,33.7,28.3,21.7.

[0058] Example 8

[0059] A quinazoline derivative, the structure of which is shown in Figure 4h; its preparation method includes the following steps:

[0060] 2 g (279.0 mg, 1.0 mmol, 1.0 eq) of the compound was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 2,7-dichloroquinazoline (199.0 mg, 1.0 mmol, 1.0 eq) was added. 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 4h (119.1 mg, yield 27%). 1 HNMR(400MHz,Chloroform-d)δ9.40(d,J=1.8Hz,1H),8.41(t,J=2.3Hz,1H),8.17(dd,J=9.2,2.0Hz,1H),8.10(dd,J=8.8,2.2Hz,1H),7.87 –7.81(m,2H),7.63(d,J=8.6Hz,1H),7.50(dd,J=9.1,2.2Hz,1H),7.17(d,J=4.6Hz,1H),7.06(dd,J=6.7,2.2Hz,1H),6.73(d,J=4.4Hz,1H). 13 C NMR(100MHz,Chloroform-d)δ167.6,155.1,153.6,150.3,136.8,135.4,135.2,12 9.3,129.2,128.2,127.0,126.7,126.1,125.2,122.9,121.6,112.0,109.7,109.3.

[0061] Example 9

[0062] A quinazoline derivative, the structure of which is shown in Figure 4i; its preparation method includes the following steps:

[0063] Compound 2h (278.1 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 2,7-dichloroquinazoline (199.0 mg, 1.0 mmol, 1.0 eq) was added. 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 4i (149.6 mg, yield 34%). 1HNMR(400MHz,Chloroform-d)δ9.40(d,J=1.8Hz,1H),8.24(t,J=2.2Hz,1H),8.17(dd,J=9.2,2.0Hz,1H),7.95(dd,J=8.9,2.3Hz,1H),7.87–7.81 (m,2H),7.64(d,J=8.9Hz,1H),7.50(dd,J=9.1,2.2Hz,1H),7.06(dd,J=6 .7,2.2Hz,1H),3.19–3.13(m,4H),1.68–1.59(m,4H),1.57–1.47(m,3H). 13 C NMR(100MHz,Chloroform-d)δ155.1,153.6,150.3,137.6,135.4,134.8,129.3,1 28.2,127.0,126.8,126.4,125.2,123.3,121.6,112.0,110.3,48.3,29.0,26.1.

[0064] Example 10

[0065] A quinazoline derivative, the structure of which is shown in Figure 4j; its preparation method includes the following steps:

[0066] Compound 2i (371.1 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 2,7-dichloroquinazoline (199.0 mg, 1.0 mmol, 1.0 eq) was added. 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 4j (149.2 mg, yield 28%). 1 HNMR(400MHz,Chloroform-d)δ9.63(s,1H),9.40(d,J=1.7Hz,1H),8.40(t,J=2.2Hz,1H),8.17(dd,J=9.2,2.1Hz,1H),8.04(dd,J=8.7,2.3Hz ,1H),7.87–7.81(m,2H),7.78(s,1H),7.64(d,J=8.6Hz,1H),7.60–7.55(m,2H),7.50(dd,J=9.1,2.3Hz,1H),7.09–7.02(m,3H),1.15(s,9H). 13C NMR(100MHz,Chloroform-d)δ177.2,155.1,153.6,150.3,136.8,135.5,135.4,134.8,133.9,1 29.3,128.2,127.0,126.7,126.2,125.2,123.2,122.7,122.6,121.6,112.0,109.7,39.4,27.6.

[0067] Example 11

[0068] A quinazoline derivative, the structure of which is shown in Figure 4k; its preparation method includes the following steps:

[0069] Compound 2a (236.1 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 2-chloroquinazoline (164.6 mg, 1.0 mmol, 1.0 eq) was added. 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 4k (134.7 mg, yield 37%). 1 H NMR(400MHz,Chloroform-d)δ9.40(d,J=1.9Hz,1H),8.25(t,J=2.2Hz,1H),8.01( dd,J=8.7,2.3Hz,1H),7.94(dt,J=8.4,1.5Hz,2H),7.83(d,J=6.7Hz,1H),7.71(t d,J=7.9,1.1Hz,1H),7.59(d,J=8.7Hz,1H),7.50–7.43(m,1H),7.11(d,J=9.1Hz, 1H), 7.06 (dd, J=6.8, 2.2Hz, 1H), 2.61 (dp, J=9.2, 5.7Hz, 1H), 0.71–0.55 (m, 4H). 13 C NMR(100MHz,Chloroform-d)δ155.6,153.8,149.4,137.1,136.0,130.9,128. 2,128.0,127.0,127.0,126.7,126.5,123.1,122.7,112.0,109.5,26.3,8.7.

[0070] Example 12

[0071] A quinazoline derivative, the structure of which is shown in Figure 41; its preparation method includes the following steps:

[0072] Compound 2a (236.1 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 7-bromo-2-chloroquinazoline (243.5 mg, 1.0 mmol, 1.0 eq) was added. 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 4l (145.9 mg, yield 33%). 1 HNMR(400MHz,Chloroform-d)δ9.45(d,J=1.8Hz,1H),8.25(t,J=2.2Hz,1H),8.21(dd,J=9.3,1.8Hz,1H),8.04–7.98(m,2H),7.83(d,J=6.7Hz,1H),7 .66(dd,J=9.2,2.2Hz,1H),7.59(d,J=8.7Hz,1H),7.11(d,J=9.1Hz,1H),7 .06(dd,J=6.8,2.2Hz,1H),2.61(dp,J=9.2,5.7Hz,1H),0.71–0.55(m,4H). 13 C NMR(100MHz,Chloroform-d)δ155.6,153.3,149.7,137.1,136.0,129.2,128. 9,128.2,127.5,126.7,126.5,124.2,122.7,121.5,112.0,109.5,26.3,8.7.

[0073] Example 13

[0074] A quinazoline derivative, the structure of which is shown in Figure 4m; its preparation method includes the following steps:

[0075] Compound 2a (236.1 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 2-chloro-7-fluoroquinazoline (182.6 mg, 1.0 mmol, 1.0 eq) was added. 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 4m (118.4 mg, yield 31%). 1HNMR(400MHz,Chloroform-d)δ9.40(d,J=1.8Hz,1H),8.25(t,J=2.1Hz,1H),8.16( ddd,J=9.1,4.9,1.9Hz,1H),8.01(dd,J=8.7,2.3Hz,1H),7.83(d,J=6.7Hz,1H),7.5 9(d,J=8.8Hz,1H),7.54(dd,J=8.1,2.2Hz,1H),7.26–7.19(m,1H),7.11(d,J=9.1H z,1H),7.06(dd,J=6.8,2.2Hz,1H),2.61(dp,J=9.2,5.7Hz,1H),0.71–0.55(m,4H). 13 C NMR(100MHz,Chloroform-d)δ163.8,161.8,155.2,153.1,151.8,151.8,137.2,136.0,128.8,128. 8,128.2,126.7,126.5,122.7,120.9,120.9,114.8,114.7,112.0,111.0,110.9,109.5,26.3,8.7.

[0076] Example 14

[0077] A quinazoline derivative, the structure of which is shown in Figure 4n; its preparation method includes the following steps:

[0078] Compound 2a (236.1 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, sodium hydride (80.0 mg, 2.0 mmol, 2.0 eq) was added in portions at 0 °C. After stirring for half an hour, 2-chloro-7-methoxyquinazoline (194.6 mg, 1.0 mmol, 1.0 eq) was added. 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 4n (141.8 mg, yield 36%). 1HNMR(400MHz,Chloroform-d)δ9.37(d,J=2.3Hz,1H),8.25(t,J=2.2Hz,1H),8.01(dd,J=8.7,2.3Hz,1H),7.86–7.77(m,2H),7.59(d,J=8.8Hz,1H),7.26(d, J=2.6Hz,1H),7.11(d,J=9.2Hz,1H),7.06(dd,J=6.7,2.2Hz,1H),6.99(dd,J= 9.1, 2.2Hz, 1H), 3.82 (s, 3H), 2.61 (dp, J=9.2, 5.7Hz, 1H), 0.71–0.55 (m, 4H). 13 C NMR(100MHz,Chloroform-d)δ160.5,156.1,152.6,150.3,137.2,136.0,128.8, 128.2,126.7,126.5,122.7,120.4,114.8,112.0,109.5,106.4,55.1,26.3,8.7.

[0079] Compound performance testing

[0080] (1) LSD1 kinase inhibition rate test

[0081] 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, 10 μL of fluorescent substrate, and 10 μL of DMSO 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 signals (excitation wavelength: 530-540 nm, emission wavelength: 585-595 nm) were acquired using a TECAN SPARK multi-functional microplate reader. The inhibition rate of the compounds on enzyme activity was then calculated. The experimental results are shown in Table 1. Compound 4b exhibited good inhibitory activity against LSD1.

[0082] Table 1. Determination of the inhibitory activity of the compounds against LSD1.

[0083] compound <![CDATA[IC 50 (nM)]]> 4a 55.1±8.2 4b 4.1±1.2 4c 120.6±5.1 4d 380.3±4.8 4e 85.8±4.2 4f 420.2±8.3 4g 18.2±3.5 4h 350.3±8.6 4i 220.4±5.7 4j 420.5±5.2 4k 45.8±6.8 41 530.9±4.4 4m 320.6±8.0 4n 150.5±9.2

[0084] (2) Determination of the cell activity of the compound

[0085] The in vitro antiproliferative activity of compound 4b 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 a CCK-8 assay, and data were processed using GraphPad Prism 9 to calculate the IC50. 50 Values. The results are shown in Table 2. Compound 4b showed good activity in the colorectal cancer cell lines HCT-116 and SW480.

[0086] Table 2 Screening of LSD1 inhibitors for cell antiproliferation activity

[0087]

[0088] (3) Compound animal tumor inhibition experiment

[0089] To evaluate the in vivo antitumor activity of the active compound, 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 mouse model was first established to evaluate the antitumor effect of compound 4b 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 examined daily to determine tumor development. When the tumors reached a certain size, mice were randomly divided into a control group and a treatment group (n=5 per group). The treatment group received an intraperitoneal injection of 50 mpk. Tumor diameter was measured daily using calipers, and mouse weight was recorded. Tumor tissue was harvested after 21 days. Changes in tumor volume were used to evaluate the antitumor effect of the compound. The results are shown in Table 3. All compounds 4b exhibited excellent in vivo antiproliferative activity.

[0090] Table 3. In vivo inhibition rate experiment

[0091]

[0092] The above description is merely a preferred embodiment of this application. This application is not limited to the above-described embodiments. Any embodiment that achieves the technical effect of this application using the same means should fall within the protection scope of this application. Within the protection scope of this application, the technical solutions and / or implementation methods can have various modifications and variations.

Claims

1. A quinazoline derivative, characterized in that, Its structure is shown in Equation I. Among them, R 1 Selected from hydrogen, cyclopropyl, methyl, phenyl, N-methylpiperazine, 4-methylhexahydropyridine, thiazole; R 2 Selected from hydrogen, chlorine, bromine, fluorine, and methoxy.

2. The quinazoline derivative according to claim 1, characterized in that, Its structure can be any one of 4a-4n.

3. The quinazoline derivative according to claim 2, characterized in that, Its structure is shown in 4b.

4. A method for preparing the quinazoline derivative according to any one of claims 1 to 3, characterized in that, The following synthetic routes are included:

5. An LSD1 inhibitor, characterized in that, Includes the quinazoline derivatives as described in any one of claims 1 to 3, or pharmaceutically acceptable salts thereof.

6. A pharmaceutical composition, characterized in that, Includes the quinazoline derivatives as described in any one of claims 1 to 3, or pharmaceutically acceptable salts thereof.

7. The pharmaceutical composition according to claim 6, characterized in that, The pharmaceutical composition includes excipients.

8. The pharmaceutical composition according to claim 7, characterized in that, The excipient is at least one of gum arabic, syrup, lanolin, and starch.

9. The use of a quinazoline derivative according to any one of claims 1 to 3 in the preparation of a reagent for diagnosing and / or treating diseases characterized by LSD1 overexpression.

10. The application according to claim 9, characterized in that, The disease in question is colorectal cancer.

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