Preparation and use of quinazoline compound

Abstract

The present invention relates to a dual-targeting EGFR T790M / L858R The invention relates to the preparation and application of quinazoline compounds of ACK1 and EGFR, which belong to the technical field of anti-tumor medicine. The technical problem solved by the invention is to provide a dual-targeting EGFR T790M / L858R and ACK1 inhibitor compounds. The compounds include the compounds shown below or their pharmaceutically acceptable salts. The compounds of the present invention or their pharmaceutically acceptable salts can be used as EGFR T790M / L858R and ACK1 inhibitors, have anti-tumor activity, and can effectively inhibit the growth of tumor cells. The compounds of the present invention are preferably anti-tumor compounds, and the corresponding tumor types are characterized by EGFR T790M / L858R and tumors with overactivation of ACK1.

Description

Technical Field

[0001] The present invention relates to a dual-targeting EGFR T790M / L858R The invention relates to the preparation and application of quinazoline compounds of the invention and ACK1, belonging to the technical field of anti-tumor medicine. Background Art

[0002] Cancer is one of the major chronic diseases that affects health, severely impacting human health and well-being and placing a heavy burden on society and families. Lung cancer is a malignant tumor originating from the bronchial mucosa or glands of the lungs. It is one of the malignant tumors with the fastest growing incidence and mortality rates, and poses the greatest threat to human health and life. Non-small cell lung cancer (NSCLC) accounts for over 80% of all lung cancers, and the 5-year survival rate of NSCLC patients is relatively low. Most patients will develop locally advanced disease or other metastatic disease. In the past decade, scientists have made significant progress in the study of different molecular and genetic subgroups of NSCLC, which has greatly changed the clinical evaluation and treatment of patients.

[0003] The epidermal growth factor receptor (EGFR) is a member of the HER family. EGFR is the expression product of the proto-oncogene c-erbB-1 and is a transmembrane tyrosine kinase receptor. EGFR plays an important role in physiological processes such as cell growth, proliferation, and differentiation by affecting downstream signaling pathways such as PI3K / AKT. Loss of EGFR function or abnormal activity can cause tumors, diabetes, immunodeficiency, and cardiovascular diseases. EGFR has also been identified as a key target for non-small cell lung cancer. Third-generation inhibitors with the advantage of combining EGFR sensitive mutations and T790M mutation sites have been successfully marketed, but the emergence of drug resistance makes it difficult for them to exert their ideal anti-tumor effects.

[0004] Studies have shown that ACK1 will be phosphorylated and activated after the third-generation EGFR inhibitors become resistant, further bypassing the activation of AKT, a downstream substrate of EGFR, thereby saving the proliferation of tumor cells. T790M / L858R Quinazoline inhibitors of ACK1 and EGFR. L858R / T790M , ACK1 and downstream signaling pathways, and showed good anti-proliferative activity in corresponding tumor cells. T790M / L858R -ACK1 inhibitors provide a potential strategy to overcome drug resistance in the treatment of non-small cell lung cancer and have broad research prospects. Summary of the Invention

[0005] The technical problem solved by the present invention is to provide a dual-targeting EGFRT790M / L858R and novel inhibitors of ACK1.

[0006] The present invention provides the following compounds or pharmaceutically acceptable salts thereof:

[0007]

[0008] Among them, R1 is

[0009] R2 is

[0010] R3 is

[0011] R4 is H, CH3, OCH3, F, Br, I, CF3, Cl.

[0012] The present invention also provides use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of anti-tumor drugs.

[0013] Furthermore, the anti-tumor drug is preferably a dual-targeting EGFR T790M / L858R and ACK1 inhibitors.

[0014] The anti-tumor drug is preferably an anti-tumor drug, and the corresponding tumor is EGFR T790M / L858R and tumors characterized by ACK1 overactivation.

[0015] The compound prepared by the present invention or its pharmaceutically acceptable salt can be used as a dual-targeting EGFR T790M / L858R and ACK1 inhibitors, have certain anti-tumor activity and can effectively inhibit the growth of cancer cells. The compounds of the present invention have a certain anti-tumor activity against various tumor cells, especially EGFR T790M / L858R The ACK1 and ACK1 overactivation pathways in non-small cell lung cancer cell lines showed a significant inhibitory effect.

[0016] Chart Description

[0017] Table 1 shows the EGFR of the above compounds. T790M / L858R and ACK1 enzyme activity detection and its antiproliferative activity test results on H1975, MCF-7, and the third-generation EGFR inhibitor osimertinib-resistant cell line AZDR.

[0018] Table 1. In vitro enzyme activity assay and antiproliferative activity results of compounds related to Formula I

[0019]

[0020] Figure 1It is the change of tumor volume in the nude mouse AZDR cell xenograft model after treatment with compound 1 in the example of the present invention.

[0021] Specific implementation party

[0022] The present invention provides the following compounds or pharmaceutically acceptable salts thereof:

[0023]

[0024] Among them, R1 is

[0025]

[0026] R2 is

[0027] R3 is

[0028] R4 is H, CH3, OCH3, F, Br, I, CF3, Cl.

[0029] Below are some preferred structures of the compounds of the present invention.

[0030]

[0031] The present invention also provides use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of anti-tumor drugs.

[0032] Furthermore, the anti-tumor drug is preferably a dual-targeting EGFR T790M / L858R and ACK1 inhibitors.

[0033] The anti-tumor drug is preferably an anti-tumor drug, and the corresponding tumor is EGFR T790M / L858R and tumors characterized by ACK1 overactivation.

[0034] The present invention also provides a pharmaceutical composition, which is a preparation containing an effective dose of the above compound or a pharmaceutically acceptable salt thereof.

[0035] The compounds of the present invention can be formulated into the following forms by methods known in the art: tablets, capsules, aqueous or oily solutions, suspensions, emulsions, creams, ointments, gels, nasal sprays, suppositories, finely divided powders for inhalation or aerosols or sprays, sterile aqueous or oily solutions or suspensions, or sterile emulsions for parenteral administration (including intravenous, intramuscular, or infusion). Liquid preparations can be prepared using sterile water or water-propylene glycol solutions as solvents, and the active ingredient can also be formulated in aqueous polyethylene glycol solutions. Aqueous solutions for oral administration can be prepared by dissolving the active ingredient in water and adding appropriate colorants, flavorings, stabilizers, and thickeners as needed. Aqueous suspensions for oral administration can be prepared by dispersing the finely divided active ingredient in water along with a viscous substance, such as natural or synthetic gums, resins, methylcellulose, carboxymethylcellulose, and other suspending agents known in the pharmaceutical art.

[0036] The pharmaceutical composition can be in unit dosage form. In these forms, the composition is divided into unit doses containing appropriate quantities of the active ingredient. The unit dosage form can be a packaged preparation containing discrete quantities of the preparation, such as boxed tablets, capsules, and powders in vials or ampoules. The unit dosage form can also be a capsule, cachet, or tablet, or any of these packaged forms can be an appropriate number.

[0037] The active ingredient of the pharmaceutical composition of the present invention may be the compound of the present invention alone, or it may be combined with other anti-tumor compounds as the active ingredient.

[0038] In the treatment of tumors, the pharmaceutical composition of the present invention can be used in combination with other anti-tumor drugs, for example, anti-proliferative / antitumor drugs, cell growth inhibitors, anti-invasion drugs, growth factor function inhibitors, anti-angiogenic agents, vascular damage agents, etc. used in medical oncology.

[0039] In the treatment of tumors, such combination therapy can be achieved by administering the various therapeutic components simultaneously, sequentially or separately. Such combination products utilize the compounds of this invention within their effective dosage ranges and the other pharmaceutically active agents within their approved dosage ranges.

[0040] The specific embodiments of the present invention are further described below in conjunction with examples, but the present invention is not limited to the scope of the examples.

[0041] Example 1 Synthesis of Preferred Compounds

[0042] The preferred compound is synthesized using the following reaction formula:

[0043]

[0044] (a) m-nitroaniline, DIPEA, isopropanol, 80℃; (b) N2H4·H2O, Renay-Ni, methanol, 0℃; (c) acryloyl chloride, TEA, THF, 0℃; (d) amino derivatives, TFA, sec-butanol, 80℃.

[0045] 1. General Synthesis of Intermediates 1-3

[0046] 2,4,6-Trichloroquinazoline (10.0 mmol) was dissolved in isopropanol (100 mL), and then 3-nitroaniline (1.11 g, 8.0 mmol) and diisopropylethylamine (6.61 mL, 40.0 mmol) were added and heated to 80°C for reaction. When TLC showed that the reaction of 3-nitroaniline was complete, the mixture was filtered under reduced pressure and the filter cake was purified by column chromatography (petroleum ether / ethyl acetate, 2 / 1) to obtain intermediate 1.

[0047] Intermediate 1 (4.0 mmol), hydrazine hydrate (497 μL, 16.0 mmol), and Raney nickel (0.47 g, 8.0 mmol) were added to methanol and reacted in an ice bath. When TLC detection showed that the reaction of intermediate 1 was complete, the mixture was filtered under reduced pressure, concentrated, and purified by column chromatography to obtain intermediate 2.

[0048] Acryloyl chloride (304 μL, 4.0 mmol) and TEA (834 μL, 6.0 mmol) were added to a solution of Intermediate 2 (2.0 mmol) in tetrahydrofuran (15 mL). The reaction was stirred in an ice bath. TLC analysis showed that the reaction of Intermediate 2 was complete. The mixture was concentrated under reduced pressure and separated by column chromatography (petroleum ether / ethyl acetate, 2 / 1) to obtain Intermediate 3.

[0049] 2. General Synthesis of Compounds 1-10

[0050] Intermediate 3 (0.5 mmol), 4-(1H-pyrazol-1-yl)aniline (0.06 g, 0.4 mmol) and TFA (111 μL, 1.5 mmol) were added to sec-butanol and stirred at 80°C. The reaction was monitored by TLC. When the reaction of 3 was complete, the target compound 1-10 was obtained by column chromatography as a yellow solid.

[0051] Compound 1, yellow solid, yield 44%; 1H NMR (400MHz, DMSO-d6) δ10.62 (d, J=196.7Hz, 3H), 8.61 (s, 1H), 7.94 (d, J=79.7Hz, 2H), 7.75 (s, 1H), 7.56 (s, 2H), 7.46 (s, 2H), 7.34 ( s, 1H), 6.48 (dd, J=16.9, 10.2Hz, 1H), 6.28 (d, J=19.0Hz, 1H), 5.78 (d, J=12.0Hz, 1H), 4.00 (d, J=91.3Hz, 2H), 1.29 (d, J=52.2Hz, 3H). 13 C NMR (100MHz, DMSO) δ163.8, 159.4, 159.2, 150.0, 135.5, 134.8, 132.3, 131.3, 129.3, 127.7, 121.0, 46.9, 16.0; HR-ESI-MS[M+H]+: m / z434.1491.

[0052] Compound 2, yellow solid, yield 35%; 1 H NMR (400MHz, DMSO-d6) δ10.2-10.0 (m, 1H), 9.3 (s, 1H), 9.1 (s, 1H), 8.5 (s, 1H), 7.9 (s, 1H), 7.6 (d, J=8.9Hz, 2H), 7.4 (s, 3H), 7.3 (t, J=8.0Hz, 1H), 6.9 (s, 1H), 6.5-6.4 (m, 1H), 6.3-6.2 (m, 1H), 5.7 (dd, J=18.9, 11.1Hz, 1H), 1.5 (s, 9H). 13 C NMR (100MHz, DMSO) δ 163.6, 163.5, 139.7, 133.6, 132.4, 129.9, 129.3, 127.4, 123.6, 123.0, 116.6, 67.6, 29.9; HR-ESI-MS [M+H]+: m / z 462.1804.

[0053] Compound 3, light yellow solid, yield 31%; 1H NMR (400MHz, DMSO-d6) δ10.20 (s, 1H), 9.67 (s, 1H), 9.11 (s, 1H), 8.56 (d, J = 31.9Hz, 1H), 8.16 (s, 2H), 7.80 (d, J = 15.0Hz, 1H), 7.68-7.41 (m, 4H ), 7.36 (s, 1H), 6.49 (dd, J=17.0, 10.1Hz, 1H), 6.28 (d, J=17.0Hz, 1H), 5.78 (d, J=12.1Hz, 1H), 3.73 (dq, J=52.4, 7.1, 6.3Hz, 4H), 3.23 (s, 3H). 13 C NMR (100MHz, DMSO) δ163.7, 140.2, 135.3, 133.6, 132.5, 130.4, 129.2, 127.2, 123.9 , 123.0, 120.6, 118.0, 116.3, 107.5, 97.6, 71.2, 58.4, 51.6; HR-ESI-MS[M+H]+: m / z 464.1596.

[0054] Compound 4, yellow solid, yield 34%; 1 H NMR (400MHz, DMSO-d6) δ10.19 (s, 1H), 9.67 (s, 1H), 9.22 (s, 1H), 8.54 (s, 1H), 8.35 (s, 1H), 8.07 (s, 1H), 7.72 (s, 1H), 7.67 (d, J = 8.9Hz, 1H) , 7.42 (d, J=27.6Hz, 3H), 7.36 (d, J=8.0Hz, 1H), 6.49 (dd, J=16.9, 10.1Hz, 1H), 6.29 (d, J=16.9Hz, 1H), 5.78 (d, J=12.1Hz, 1H), 5.08 (s, 2H). 13 C NMR (100MHz, DMSO-d6) δ163.7, 151.3, 140.0, 139.7, 133.6, 132.5, 132.4, 129.3, 127.4, 125.0, 123.0, 121.5, 51.9; HR-ESI-MS[M+H]+: m / z 488.1208.

[0055] Compound 5, yellow solid, yield 29%; 1H NMR (400MHz, DMSO-d6) δ10.26 (s, 1H), 9.89-8.74 (m, 2H), 8.57 (s, 1H), 7.98 (d, J=114.8Hz, 3H), 7.70 (s, 3H), 7.48 (s, 4H), 7.37 (t, J=8.OHz, 2H), 7.27 (t, J=7.5Hz, 1H), 6.46 (s, 1H), 6.26 (d, J=16.9Hz, 1H), 5.76 (d, J=4.9Hz, 1H). 13 C NMR (100MHz, DMSO) δ163.7, 156.0, 140.2, 139.8, 133.8, 1323, 129.9, 127.4, 126.3, 126.0, 123.0, 118.1, 116.8. HPLC purity: 95.84%; HR-ESI-MS[M+H]+: m / z482.1491.

[0056] Compound 6, yellow solid, yield 33%; 1 H NMR (400MHz, DMSO-d6) δ10.19 (s, 1H), 9.62 (s, 1H), 9.15 (s, 1H), 8.52 (s, 1H), 8.18 (d, J = 77.1Hz, 2H), 7.78 (s, 1H), 7.68-7 .41 (m, 4H), 7.38-7.17 (m, 6H), 6.48 (dd, J=16.9, 10.2Hz, 1H), 6.29 (d, J=16.9Hz, 1H), 5.77 (d, J=12.2Hz, 1H), 5.30 (s, 2H). 13 C NMR (100 MHz, DMSO-d 6 )δ163.7, 132.4, 130.7, 129.3, 128.9, 127.9, 127.9, 127.4, 124.3, 123.0, 120.3, 55.4; HR-ESI-MS[M+H]+: m / z 496.1647.

[0057] Compound 7, yellow solid, yield 28%; 1H NMR (400MHz, DMSO-d6) δ10.2 (s, 1H), 9.6 (s, 1H), 9.1 (s, 1H), 8.5 (s, 1H), 8.2 (d, J = 35.6Hz, 1H), 7.8 (d, J = 78.8Hz, 1H), 7.6 (d, J = 8.9Hz, 3H), 7.4 (t, J =10.0Hz, 2H), 7.2 (s, 1H), 6.5 (dd, J = 16.9, 10.1Hz, 1H), 6.3 (d, J = 16.9Hz, 1H), 5.8 (d, J=12.1Hz, 1H), 4.4 (s, 1H), 3.9 (s, 2H), 3.4 (s, 2H), 1.9 (s, 4H). 13 C NMR (100MHz, DMSO) δ163.7, 157.9, 139.7, 133.9, 133.6, 132.4, 130.1, 127. 5, 123.6, 123.0, 118.3, 66.4, 57.6, 33.4, 31.7, 30.3; HR-ESI-MS[M+H]+: m / z 490.1753.

[0058] Compound 8, yellow solid, yield 31%; 1 H NMR (400 MHz, DMSO-d 6 )δ10.3 (s, 1H), 9.6 (s, 1H), 9.2 (s, 1H), 8.5 (s, 1H), 8.2 (s, 1H), 7.8-7.6 (m, 3H), 7.5 (s, 2H), 7.4 (s, 2H), 6.5 (t, J=1 1.6Hz, 1H), 6.3 (d, J=19.0Hz, 1H), 5.8 (d, J=12.2Hz, 1H), 4.1 (s, 1H), 3.0 (s, 3H), 2.3 (s, 4H), 1.9 (d, J=85.6Hz, 4H). 13 C NMR (100MHz, DMSO) δ163.7, 158.6, 158.3, 140.2, 139.8, 139.5, 139.1, 132.5, 130.1, 127.2 , 123.7, 123.0, 119.2, 118.3, 117.9, 116.2, 57.6, 54.2, 45.4, 31.8; HR-ESI-MS[M+H]+: m / z 503.2069.

[0059] Compound 9, yellow solid, yield 27%; 1 H NMR (400 MHz, DMSO-d 6)δ10.20 (s, 1H), 9.61 (s, 1H), 9.11 (s, 1H), 8.52 (s, 1H), 8.11 (s, 1H), 7.78 (s, 1H), 7.70-7.19 (m, 6H), 6.49 (dd, J=16.9, 10 .1Hz, 1H), 6.29 (d, J=19.1Hz, 1H), 5.78 (d, J=12.1Hz, 1H), 4.07 (d, J=93.5Hz, 2H), 3.54 (s, 4H), 2.69 (s, 2H), 2.40 (s, 4H). 13 C NMR (100MHz, DMSO) δ163.7, 161.9, 133.6, 132.4, 130.2, 129.3, 127.4, 123.8, 123.0, 120.5, 66.6, 58.4, 53.6, 49.3; HR-ESI-MS[M+H]+: m / z519.2018.

[0060] Compound 10, yellow solid, yield 43%; 1 H NMR (400 MHz, DMSO-d 6 )δ12.38 (s, 1H), 10.18 (s, 1H), 9.62 (s, 1H), 9.12 (s, 1H), 8.53 (s, 1H), 8.12 (s, 1H), 7.88 (s, 1H), 7.77 (s, 1H), 7.64 (d, J=6. 6Hz, 1H), 7.45 (s, 3H), 7.34 (t, J=8.1Hz, 1H), 6.50 (dd, J=16.9, 10.1Hz, 1H), 6.28 (d, J=19.0Hz, 1H), 5.77 (d, J=12.1Hz, 1H). 13 C NMR (101MHz, DMSO) δ163.6, 157.8, 156.5, 151.2, 140.1, 139.7, 133.5, 132.4, 129.2, 127.3, 123.4, 123.0, 115.4, 114.0; HR-ESI-MS[M+H]+: m / z 406.1178.

[0061] Test Example 1 Antiproliferation test of compound 1 on NCI-H1975, MCF-7, AZDR cells

[0062] Antiproliferative activity assay in NCI-H1975, MCF-7, and AZDR cells:

[0063] NCI-H1975, MCF-7, and AZDR cells in the logarithmic growth phase were seeded in 96-well plates (approximately 5×10 cells per well). 3cells). Culture for 24 hours. After the cells adhere to the wall and grow, add the specified concentration of each compound and incubate for 24 hours. Use PBS to dissolve MTT to a final concentration of 0.5%, and then add 20 μL of MTT solution to each well. Incubate in a cell culture incubator at 37°C and 5% CO2 for 3-4 hours in the dark, then discard the existing culture medium, add 150 μL of DMSO to each well, detect the corresponding OD value at a wavelength of 490nM using an enzyme marker, analyze the effect of each compound on cell proliferation using SPSS, and evaluate the anti-proliferative activity of each compound. The results are shown in Table 1. It can be clearly seen that this series of compounds has a significant and strong anti-proliferative activity against tumor cells. Among them, the preferred compound 1 has a strong anti-proliferative activity against EGFR L858R / T790M It has significant and strong anti-proliferative activity against cells over-activated by ACK1, with the half-maximal inhibitory concentrations for NCI-H1975, MCF-7, and AZDR cells being 0.67±0.08μM, 1.04±0.15μM, and 1.27±0.12μM, respectively.

[0064] Experimental Example 2: In vivo antitumor activity experiment of compound 1

[0065] The purpose of this study was to examine the in vivo anti-tumor efficacy of the preferred compounds of this invention. Compound 1 was administered intraperitoneally in an osimertinib-resistant AZDR cell xenograft mouse model at doses of 20, 40, or 60 mg / kg once daily. The ACK1 inhibitor dasatinib, the third-generation EGFR inhibitor osimertinib, and a combination of both drugs were used as positive controls at a dose of 10 mg / kg.

[0066] The tumor growth curves measured experimentally are as follows Figure 1 As shown, oral administration of osimertinib and dasatinib induced a certain degree of tumor regression. Compound 1 exerted significant in vivo anti-tumor activity in a dose-dependent manner. At 60 mg / kg, compound 1 showed superior in vivo anti-tumor activity compared to the combination of dasatinib (10 mg / kg) and osimertinib (10 mg / kg).

[0067] Combining the above experiments, we obtained the ability to effectively inhibit EGFR L858R / T790M and ACK1 to exert anti-cancer effects, and such inhibitors can be used to treat EGFR L858R / T790 Tumors with overactivation of M and ACK1 provide a good approach for the development of such inhibitors and have broad research prospects.

Claims

1. Compounds having the structural formulae I to X or pharmaceutically acceptable salts thereof:

2. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of anti-tumor drugs.

3. The use according to claim 2, characterized in that: The anti-tumor drug is a dual-targeting EGFR T790M / L858R and ACK1 inhibitors.

4. The use according to claim 2, characterized in that: The anti-tumor drug is for treating EGFR T790M / L858R and drugs for tumor types characterized by ACK1 overactivation.

5. A pharmaceutical composition, characterized in that: The invention relates to a preparation comprising an effective dose of the compound according to claim 1 or a pharmaceutically acceptable salt thereof.

Images