A quinoline compound with dual inhibitory effects on aurora kinase B and epidermal growth factor receptor and its use

Abstract

The present invention discloses a quinoline compound having a general structural formula selected from any one of the following: #imgabs0#, #imgabs1#, etc. The present invention also provides the use of the compound in preparing a drug having dual inhibitory effects on Aurora kinase B and epidermal growth factor receptor. The drug prepared using the compound has anti-non-small cell lung cancer effects.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine and relates to a class of quinoline compounds having dual inhibitory effects on aurora kinase B and epidermal growth factor receptor and uses thereof. Background Art

[0002] Cancer, a major global public health issue, severely impacts social development. The primary goal of cancer treatment is to inhibit the migration and proliferation of malignant tumor cells. Clinical approaches such as chemotherapy, targeted drug therapy, and immunotherapy are widely used.

[0003] Epidermal growth factor receptor (EGFR) inhibitors have demonstrated promising clinical efficacy, particularly in the treatment of cancers such as non-small cell lung cancer (NSCLC). For patients harboring EGFR mutations (such as L858R and Exon 19 deletion mutations), initial treatment with EGFR inhibitors (such as gefitinib and erlotinib) typically results in a 50-70% response rate and a progression-free survival of up to nine months for patients with NSCLC harboring EGFR mutations. Although EGFR inhibitors are effective in the early stages of treatment, patients develop acquired resistance after 9-13 months of treatment, with the EGFR T790M mutation being the most common mechanism of resistance. The development of inhibitors specific for these resistance targets is a key approach to improving survival in patients with malignant tumors. Osimertinib, a third-generation EGFR inhibitor designed to target the EGFR T790M mutation, has an acrylamide group that forms an irreversible covalent bond with the cysteine ​​residue C797 in the ATP-binding pocket of the EGFR T790M mutation. Osimertinib's molecular backbone is highly flexible, adapting to the steric constraints of the ATP-binding pocket caused by the T790M mutation. Consequently, osimertinib has a response rate of 80% and a progression-free survival of 18.9 months. Unfortunately, some patients develop other resistance mechanisms (such as the C797S mutation) after treatment, reducing the therapeutic effect.

[0004] Aurora kinase B (Aurora-B) is a serine-threonine kinase that is primarily involved in chromosome cohesion, segregation, and cytokinesis. During cell division, Aurora-B is localized to the metaphase spindle and helps separate the two daughter cells at the completion of cytokinesis. Due to its importance for chromosome stability and cell division, aberrant activation or overexpression often leads to alterations in chromosome number and structure, causing cells to lose control during mitosis. This chromosomal instability often leads to malignant transformation. Aneuploidy, meaning an abnormal number of chromosomes, is found in 90% of human malignant tumors. This may be caused by defects in mitotic checkpoints and chromosome cohesion, which have been linked to Aurora-B overexpression. Therefore, Aurora-B has been established as a target for cancer therapy. Studies have shown that knocking down Aurora-B expression inhibits activation of the mTOR / ULK1 pathway, stimulates autophagy in osteosarcoma cells, and suppresses metastasis and proliferation. Further studies have found that in drug-resistant tumor cells, Aurora-B promotes the development of drug resistance in malignant tumor cells through multiple pathways: 1. The CDK7-Aurora-B pathway is upregulated in two types of AML cells that are resistant to FTL3 inhibitors, and Aurora-B and cyclin-dependent kinases 2 / 4 / 6 work together to slow down cell metabolic activity, thereby enhancing adaptability to FTL3 inhibitors; 2. Aurora-B and MYC Mutual activation enhances the proliferation of malignant tumors; 3. Aurora-B inhibits apoptosis in malignant tumor cells through the AKT / PI3K pathway; and in the aforementioned study, combined use of an Aurora-B inhibitor restored tumor cell sensitivity to the drug, effectively improving drug resistance. Given the multifaceted connection between Aurora-B and drug resistance, the development of multi-targeted small molecule drugs, including those targeting Aurora-B, offers new possibilities for improving tumor drug resistance.

[0005] To date, corresponding drug resistance mechanisms have been found in clinical practice for first- to third-generation single-target EGFR inhibitors. Inhibition of only a single pathway can promote the activation of related pathways, suggesting that when treating tumors with complex pathogenic mechanisms, a strategy that simultaneously inhibits multiple targets is more appropriate. Literature research revealed that the EGFR inhibitor pelitinib, the Aurora-B inhibitor AL8326, and anlotinib all use quinoline as their core structure. Therefore, this study used quinoline as the molecular backbone and utilized computer-aided drug design to rationally design and modify quinoline substituents to construct a quinoline-based Aurora-B / EGFR dual-target inhibitor.

[0006] Therefore, the object of the present invention is to provide quinoline-derived dual inhibitors of Aurora-B / EGFR with improved selectivity for osimertinib-resistant non-small cell lung cancer malignancies. Summary of the Invention

[0007] In response to the above-mentioned technical problems in the prior art, the present invention provides a quinoline compound with dual inhibitory effects on Aurora kinase B and epidermal growth factor receptor and its use. The quinoline compound with dual inhibitory effects on Aurora kinase B and epidermal growth factor receptor and its use are intended to solve the problem of osimertinib resistance in patients with EGFR mutation NSCLC in the prior art.

[0008] The present invention provides a quinoline compound selected from any one of the following structural formulas:

[0009] 、 、 、 、 、 、 、 .

[0010] The present invention also provides pharmaceutically acceptable salts, solvates, polymorphs, isomers or prodrugs of the above-mentioned quinoline compounds.

[0011] The present invention also provides a pharmaceutical composition comprising the above compound and a pharmaceutically acceptable carrier and / or excipient.

[0012] The present invention also provides a pharmaceutical composition comprising the above-mentioned pharmaceutically acceptable salt, solvate, polymorph, isomer or prodrug and a pharmaceutically acceptable carrier and / or excipient.

[0013] Furthermore, the pharmaceutically acceptable adjuvant and / or excipient is a diluent, a binder, a surfactant, a wetting agent, an adsorption carrier, a lubricant, a filler and / or a disintegrant.

[0014] Furthermore, the dosage form of the drug is an aqueous dispersion, liquid, gel, syrup, slurry, suspension, aerosol, controlled release agent, fast dissolving agent, effervescent agent, lyophilized agent, tablet, powder, pill, dragee, capsule, delayed release agent, extended release agent, pulse controlled release agent, multi-particulate or immediate release agent.

[0015] The present invention also provides the use of the compound in preparing a drug having dual inhibitory effects on aurora kinase B and epidermal growth factor receptor.

[0016] The present invention also provides the use of the pharmaceutically acceptable salt, solvate, polymorph, isomer or prodrug in the preparation of a drug having dual inhibitory effects on Aurora kinase B and epidermal growth factor receptor.

[0017] The present invention also provides the use of the above-mentioned pharmaceutically acceptable compound in the preparation of a drug for treating tumors associated with overexpression of Aurora kinase B and epidermal growth factor receptor

[0018] The present invention also provides use of the pharmaceutically acceptable salt, solvate, polymorph, isomer or prodrug in the preparation of a medicament for treating tumors associated with overexpression of Aurora kinase B and epidermal growth factor receptor.

[0019] The present invention also provides use of the compound in preparing medicine for treating non-small cell lung cancer.

[0020] The present invention also provides use of the pharmaceutically acceptable salt, solvate, polymorph, isomer or prodrug in preparing a drug for treating non-small cell lung cancer.

[0021] Compared with the existing technology, the present invention has significant technical progress. The present invention discloses a series of Aurora-B / EGFR dual-target compounds, which provides a new direction for the research of anti-non-small cell lung cancer drugs and is of great significance for the development of anti-non-small cell lung cancer drugs.

[0022] It should be understood that within the scope of the present invention, the above-mentioned technical features and the technical features described in detail below (e.g., in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they are not described in detail here. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The molecular docking combination of the quinoline derivative disclosed in the prior art and Aurora-B is shown. DETAILED DESCRIPTION

[0024] Through extensive and intensive research, the present inventors unexpectedly discovered a series of novel quinoline compounds that exhibit potent dual inhibitory activity against EGFR and Aurora-B. This led to the present invention. The compounds described above can be synthesized using standard synthetic techniques or known techniques combined with the methods described herein. Furthermore, the solvents, temperatures, and other reaction conditions described herein may vary.

[0025] The starting materials used in the synthesis of the compounds can be synthesized or obtained from commercial sources. The compounds described herein and other related compounds having various substituents can be synthesized using known techniques and starting materials. The general methods for preparing the compounds can be modified by using appropriate reagents and conditions for introducing various groups into the molecular formulas provided herein. The compounds described herein are synthesized using the synthetic routes shown in the schemes below. In some embodiments, the compounds described herein can be prepared by the methods described below. The following methods and examples are intended to illustrate these methods. These schemes and examples should not be construed as limiting the present invention in any way. The compounds described herein can also be synthesized using standard synthetic techniques known to those skilled in the art, or a combination of methods known in the art and the methods described herein.

[0026] The synthesis method of the series of compounds may include the following steps:

[0027]

[0028] Example 1 Synthesis of Intermediate 3:

[0029]

[0030] 7-Benzyloxy-4-chloro-6-methoxyquinoline and 4-fluoro-5-hydroxy-2-methylindole were reacted in 2,6-lutidine under the catalysis of 4-dimethylaminopyridine at 140°C. After TLC detection, the reaction was complete, and the mixture was cooled to room temperature, diluted with dichloromethane, and 50 mL of 1 mol / L HCl aqueous solution was slowly added dropwise. The aqueous layer was discarded, and the organic layer was washed with 1 mol / L potassium carbonate solution, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by distillation under reduced pressure. 1.74 g of intermediate 3 was isolated by column chromatography as a light yellow solid in a yield of 51.15%. 1 H NMR (400 MHz, DMSO- d 6) δ 11.42 (t, J = 2.5 Hz, 1H), 8.41(d, J = 5.2 Hz, 1H), 7.61 (s, 1H), 7.57 – 7.50 (m, 3H), 7.50 (s, 2H), 7.43(dd, J = 8.2, 6.6 Hz, 2H), 7.40 – 7.32 (m, 1H), 7.22 (d, J = 8.6 Hz, 1H),7.04 – 6.94 (m, 1H), 6.33 (dd, J= 5.2, 1.1 Hz, 1H), 6.30 – 6.25 (m, 1H),5.31 (s, 2H), 3.97 (s, 3H), 3.32 (s, 2H), 2.42 (s, 3H).LC-MS (ESI) m / z 429.2[M+H] + .

[0031] Example 2 Synthesis of Intermediate 4:

[0032]

[0033] Synthesis route of intermediate 4

[0034] 7-Benzyloxy-4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinoline (800 mg, 1.87 mmol) and 3 mL of H₂O were added to a three-necked flask. The temperature was raised to 85°C with stirring, and 12 mL of trifluoroacetic acid was added. The mixture was stirred for 2 hours. After TLC analysis of the reaction, the mixture was cooled to room temperature. Saturated sodium bicarbonate solution was added to the reaction solution to adjust the pH to 7-8. The solution was extracted three times with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and filtered. The organic solvent was evaporated under reduced pressure. The product was purified by column chromatography to yield 550 mg of intermediate 4 as a pale yellow solid in an 87% yield. 1 H NMR (400 MHz, DMSO- d 6) δ 11.58 (t, J =2.4 Hz, 1H), 8.68 (d, J = 6.5 Hz, 1H), 7.77 (s, 1H), 7.50 (s, 1H), 7.29 (d, J = 8.6 Hz, 1H), 7.18 – 7.05 (m, 1H), 6.72 (dd, J = 6.6, 1.1 Hz, 1H), 6.36 –6.30 (m, 1H), 4.05 (s, 4H). LC-MS (ESI) m / z 339.1 [M+H] + .

[0035] Example 3 Synthesis of Intermediate 5:

[0036]

[0037] Synthesis route of intermediates 5 and 6

[0038] At room temperature, 2-bromoethylamine hydrobromide, H2O, and CH3COOH were added to a three-necked flask, followed by 2,5-dimethoxytetrahydrofuran and sodium acetate. The solution was stirred at room temperature until it became clear and then heated to 55°C. After the reaction, saturated sodium bicarbonate was added to adjust the pH of the solution to 7-8. The solution was extracted with dichloromethane three times. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain 1.10 g of intermediate 5 as a light yellow oil with a yield of 63.20%. The reaction steps for intermediate 6 were the same as those for intermediate 5, except that the column chromatography separation system (dichloromethane:methanol = 70:1) was used. 1 H NMR (400 MHz, DMSO- d 6) δ 6.81 (t, J = 2.2 Hz, 1H), 6.03 (t, J = 2.1 Hz, 1H), 4.27 (t, J = 6.4 Hz, 1H), 3.74 (t, J = 6.4 Hz, 1H). LC-MS(ESI) m / z 175.9 [M+H] + .

[0039] Example 4 Synthesis of Intermediate 6:

[0040] The synthesis was similar to the steps described in Example 3, except that 2-bromoethylamine hydrobromide was replaced by 3-bromopropylamine hydrobromide, to obtain 940 mg of light yellow oil with a yield of 54.01%. 1 H NMR (400 MHz, DMSO- d 6) δ 6.74 (t, J =2.1 Hz, 2H), 6.00 (t, J = 2.1 Hz, 2H), 3.99 (t, J = 6.7 Hz, 2H), 3.37 (t, J =6.5 Hz, 2H), 2.20 (p, J = 6.6 Hz, 2H). LC-MS (ESI) m / z 190.0 [M+H] + .

[0041] Example 5 Synthesis of Intermediate 7

[0042]

[0043] Synthesis route of intermediate 7

[0044] Add 1H-1,2,4-triazole, potassium bicarbonate, dibromopropane into a three-necked flask. N,N -dimethylformamide, react at room temperature. After the reaction, extract with 30 mL of dichloromethane three times, backwash once with water, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under reduced pressure at low temperature to obtain 1.34 g of intermediate 7 as a light yellow oil with a yield of 7.06%. 1 H NMR (600 MHz, DMSO- d 6) δ8.55 (s, 1H), 7.99 (s, 1H), 4.32 (t, J = 6.8 Hz, 2H), 3.49 (t, J = 6.5 Hz,2H), 2.33 (p, J = 6.7 Hz, 2H); LC-MS (ESI) m / z 190.1 [M+H] + .

[0045] Example 6 Synthesis of Compound 13:

[0046]

[0047] 4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-7-[2-(1H-pyrrol-1-yl)ethoxy]quinoline

[0048] To a three-necked flask, the intermediate 4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-7-hydroxyquinoline (200 mg, 591.12 μmol), 1-(2-bromoethyl)-1H-pyrrole (120 mg, 709.35 μmol), potassium carbonate (245 mg, 1.77 mmol), and 4 mL of N,N-dimethylformamide were added. The reaction was carried out at 60-80 °C. After the reaction was completed, 20 mL of water was added, and the mixture was extracted three times with 30 mL of ethyl acetate. The aqueous layer was discarded, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (ethyl acetate: cyclohexane = 2:1) 120 mg of the final product 4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-7-[2-(1H-pyrrol-1-yl)ethoxy]quinoline was isolated as a light yellow solid in a yield of 47.05%. 1HNMR (600 MHz, DMSO-d6) δ 11.42 (s, 1H), 8.42 (d, J = 5.2 Hz, 2H), 7.61 (s,2H), 7.41 (s, 2H), 7.22 (d, J = 8.5 Hz, 1H), 7.02 – 6.97 (m, 1H), 6.90 (t, J= 2.1 Hz, 3H), 6.33 (d, J = 5.2 Hz, 1H), 6.28 (d, J = 1.1 Hz, 1H), 6.01 (t, J= 2.1 Hz, 3H), 4.42 (d, J = 5.2 Hz, 4H), 4.38 (t, J = 4.7 Hz, 4H), 3.98 (s,5H), 2.42 (s, 3H). LC-MS (ESI) m / z 431.8 [M+H]+ .

[0049] Example 7 Synthesis of Compound 14:

[0050]

[0051] 4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-7-[2-(1H-imidazol-1-yl)ethoxy]quinoline

[0052] The synthesis was similar to the steps described in Example 6, except that 1-(2-bromoethyl)-1H-pyrrole was replaced by 1-(2-chloroethyl)imidazole hydrochloride, with a yield of 19.56%. 1H NMR (400 MHz, DMSO-d6) δ 11.48 (t, J = 2.4Hz, 1H), 8.43 (d, J = 5.2 Hz, 1H), 7.74 (s, 1H), 7.61 (s, 1H), 7.43 (s, 1H),7.31 (d, J = 1.4 Hz, 1H), 7.22 (d, J = 8.6 Hz, 1H), 7.04 – 6.93 (m, 1H), 6.91 (d, J = 1.2 Hz, 1H), 6.34 (dd, J = 5.3, 1.1 Hz, 1H), 6.28 (d, J = 2.0 Hz,1H), 4.51 – 4.42 (m, J = 3.0 Hz, 4H), 3.98 (s, 3H), 2.42 (s, 3H). LC-MS (ESI)m / z 433.1 [M+H]+.

[0053] Example 8 Synthesis of Compound 15:

[0054]

[0055] 4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-7-[2-(1H-pyrazol-1-yl)ethoxy]quinoline

[0056] The synthesis was similar to the steps described in Example 6, except that 1-(2-bromoethyl)-1H-pyrrole was replaced by 1-(2-bromoethyl)pyrazole, with a yield of 7.82%. 1H NMR (600 MHz, DMSO-d6) δ 11.42 (s, 1H), 8.43 (d, J =5.2 Hz, 2H), 7.84 (d, J = 2.3 Hz, 2H), 7.59 (s, 2H), 7.49 (d, J = 1.9 Hz, 2H), 7.42 (s, 2H), 7.22 (d, J = 8.6 Hz, 2H), 6.99 (t, J = 8.1 Hz, 2H), 6.33(d, J = 5.2 Hz, 1H), 6.28 (t, J = 2.1 Hz, 4H), 4.61 (t, J = 5.1 Hz, 4H), 4.54(t, J = 5.2 Hz, 4H), 3.95 (s, 6H), 2.42 (s, 3H). LC-MS (ESI) m / z 433.2 [M+H]+ .

[0057] Example 9 Synthesis of Compound 16:

[0058]

[0059] 4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-7-[2-(1H-1,2,4-triazol-1-yl)ethoxy]quinoline

[0060] The synthesis was similar to the steps described in Example 6, except that 1-(2-bromoethyl)-1H-pyrrole was replaced by 1-(2-chloroethyl)-1H-1,2,4-triazole, with a yield of 21.52%. 1H NMR (400 MHz, DMSO-d6) δ 11.42 (s, 1H), 8.60 (s, 1H), 8.43 (d, J = 5.3 Hz, 1H), 8.01 (s, 1H), 7.59 (s, 1H), 7.45 (s,1H), 7.22 (d, J = 8.6 Hz, 1H), 7.03 – 6.96 (m, 1H), 6.34 (dd, J = 5.3, 1.1Hz, 1H), 6.30 – 6.26 (m, 1H), 4.70 (t, J = 5.1 Hz, 2H), 4.56 (t, J = 5.0 Hz,2H), 3.94 (s, 3H), 2.42 (s, 3H). LC-MS (ESI) m / z 434.2 [M+H]+.

[0061] Example 10 Synthesis of Compound 17:

[0062]

[0063] 4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-7-[2-(1H-pyrrol-1-yl)propoxy]quinoline

[0064] The synthesis was similar to the steps described in Example 6, except that 1-(2-bromoethyl)-1H-pyrrole was replaced by 1-(3-bromopropyl)-1H-pyrrole, with a yield of 7.82%. 1H NMR (400 MHz, DMSO-d6) δ 11.43 (s, 1H), 8.41 (d,J = 5.3 Hz, 1H), 7.61 (s, 1H), 7.35 (s, 1H), 7.22 (d,J = 9.4 Hz, 1H), 7.02 –6.96 (m, 1H), 6.78 (t, J = 2.1 Hz, 2H), 6.33 (dd, J = 5.2, 1.1 Hz, 1H), 6.28(s, 1H), 5.99 (t, J = 2.1 Hz, 2H), 4.11 (t, J = 6.8 Hz, 2H), 4.06 (t, J = 6.2Hz, 3H), 3.99 (s, 3H), 2.42 (s, 3H), 2.25 (p, J = 6.6 Hz, 3H). LC-MS (ESI) m / z 446.2 [M+H]+ .

[0065] Example 11 Synthesis of Compound 18:

[0066]

[0067] 4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-7-[2-(1H-imidazol-1-yl)propoxy]quinoline

[0068] The synthesis was similar to the steps described in Example 6, except that 1-(2-bromoethyl)-1H-pyrrole was replaced by 1-(3-bromopropyl)-1H-imidazole hydrobromide, with a yield of 9.47%. 1H NMR (400 MHz, DMSO-d6) δ 11.42 (s, 1H), 8.42 (d, J = 5.3 Hz, 1H), 7.67 (s, 1H), 7.61 (s, 1H), 7.38 (s, 1H), 7.24 (s,1H), 7.22 (d, J = 8.6 Hz, 1H), 7.03 – 6.96 (m, 1H), 6.91 (s, 1H), 6.33 (dd, J= 5.2, 1.1 Hz, 1H), 6.31 – 6.25 (m, 1H), 4.20 (t, J = 6.9 Hz, 2H), 4.10 (t, J= 6.3 Hz, 2H), 3.99 (s, 3H), 2.42 (s, 3H), 2.29 (p, J = 6.5 Hz, 2H). LC-MS(ESI) m / z 447.2 [M+H]+ .

[0069] Example 12 Synthesis of Compound 19:

[0070]

[0071] 4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-7-[2-(1H-pyrazol-1-yl)propoxy]quinoline

[0072] The synthesis was similar to the steps described in Example 6, except that 1-(2-bromoethyl)-1H-pyrrole was replaced by 1-(3-chloropropyl)-1H-pyrazole hydrochloride, with a yield of 11.37%. 1H NMR (400 MHz, DMSO-d6) δ 11.42 (s, 1H), 8.42 (d, J = 5.3 Hz, 1H), 7.76 (d, J = 1.6 Hz, 1H), 7.61 (s, 1H), 7.46 (d, J= 1.2 Hz, 1H), 7.35 (s, 1H), 7.22 (d, J = 8.6 Hz, 1H), 7.03 – 6.96 (m, 1H), 6.33 (dd, J = 5.3, 1.1 Hz, 1H), 6.28 (s, 1H), 6.25 (t, J = 2.0 Hz, 1H), 4.35(t, J = 6.8 Hz, 2H), 4.13 (t, J = 6.2 Hz, 2H), 3.99 (s, 2H), 2.42 (s, 2H), 2.33 (p, J = 6.6 Hz, 2H). LC-MS (ESI) m / z 447.2 [M+H]+ .

[0073] Example 13 Synthesis of Compound 20:

[0074]

[0075] 4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-7-[2-(1H-1,2,4-triazol-1-yl)propoxy]quinoline

[0076] The synthesis was similar to the steps described in Example 6, except that 1-(2-bromoethyl)-1H-pyrrole was replaced by 1-(3-bromopropyl)-1H-1,2,4-triazole, with a yield of 16.26%. 1H NMR (600 MHz, DMSO-d6) δ 11.42 (s, 1H),8.57 (s, 1H), 8.42 (d, J = 5.2 Hz, 1H), 8.00 (s, 1H), 7.61 (s, 1H), 7.38 (s,1H), 7.22 (d, J = 8.5 Hz, 1H), 7.02 – 6.98 (m, 1H), 6.33 (d, J = 5.2 Hz, 1H), 6.28 (s, 1H), 4.42 (t, J = 6.9 Hz, 2H), 4.18 (t, J = 6.2 Hz, 2H), 3.98 (s,3H), 2.42 (s, 3H), 2.36 (p, J = 6.6 Hz, 2H). LC-MS (ESI) m / z 448.2 [M+H]+ .

[0077] Example 14: Determination of IC of target compounds against Aurora-B and EGFR using HTRF 50 value.

[0078] Experimental materials: EGFR was purchased from Carna, ATP, MgCl2 and MnCl2 were purchased from Sigma, and EGFR detection kit was purchased from Revvity. 50 Testing performed by BioDuro Biotech (Jiangsu) Co., Ltd.

[0079] Following the HTRF kinEASE TK / STK Kit instructions, prepare stock solutions of TK / STK-Substrate-biotin, streptavidin-XL665, TK / STK antibody-cryptate, and EGFR / Aurora-B. Dilute each kinase stock solution with 1x kinase buffer as needed to prepare working solutions at the desired concentrations for the enzymatic step. Each test compound is diluted in 1x kinase buffer to ensure a DMSO concentration of ≤2% and then diluted eight times in three-fold series to generate nine concentrations for the enzymatic step. Each working concentration is 2.5x the final concentration (range: 10 μM). Blank and control compounds are assayed in a white 96-well microplate.

[0080] The ratio of the two fluorescence intensities was calculated and the inhibition rate of each compound on the kinase at a given concentration was expressed as (Ratio 待测化合物 -RatioHigh Control ) / (Ratio Low Control - Ratio High Control ), where Ratio Low Control 、Ratio High Control The release rates of the negative control wells and blank control wells are shown in Table 1. The half-maximal inhibitory concentration (IC 50 ) were determined by dose-response curves obtained with GraphPad Prism9 software.

[0081] Table 1 IC of compounds against Aurora-B and EGFR 50

[0082]

[0083] As shown in Table 1, a total of 8 target compounds have been synthesized. After structural confirmation and purity verification, the inhibitory activities of the 8 target compounds and anlotinib against Aurora-B and EGFR were determined using homogeneous time-resolved fluorescence (HTRF) technology. For EGFR, the inhibition rates of the series of compounds were lower than those of anlotinib to varying degrees. Among them, the inhibitory activities of 14, 15, 16, 18, and 20 against Aurora-B were better than those of anlotinib, and the IC of 16 was 50 It is 0.833μM.

[0084] The quinoline derivatives disclosed in Examples 17 and 18 of the prior art (publication number CN1446212A, publication date October 1, 2003) only have VEGF inhibitory activity. The present invention provides a quinoline compound with dual inhibitory effects on Aurora kinase B and epidermal growth factor receptor.

[0085] Example 15: In this experiment, the CCK-8 method (Cell Counting Kit-8) was used to detect the in vitro cell anti-proliferative activity of the target compound.

[0086] Establishment of drug-resistant cells: The osimertinib-resistant H1975 cell line was established using a low-dose continuous induction method to ensure that the H1975 cell line can continue to grow at a concentration of 5 μM osimertinib. The resistant cell line exhibits slow proliferation, low apoptosis, and high migration and invasion capabilities.

[0087] Preparation of complete culture medium: The complete culture medium for osimertinib-resistant H1975 cells is composed of 89% basal culture medium + 10% fetal bovine serum + 1% penicillin-streptomycin dual antibody.

[0088] Dilution of compound solution: Prepare a 20 mM stock solution of the test drug. Add the required volume of dimethyl sulfoxide. To ensure complete dissolution, sonication, vortexing, or heating can be used to accelerate dissolution. Before use, dilute the solution in a clean hood with complete culture medium to concentrations of 2, 4, 6, 8, 10, or 12 μM. Mix thoroughly and set aside.

[0089] Plating: Take tumor cells in the logarithmic growth phase, digest them with trypsin, centrifuge and remove the old culture medium; add 1 mL of complete culture medium to suspend. Take 10 μL of cell suspension to a new EP tube, add 90 μL of complete culture medium to dilute, transfer to a cell counting plate, count, and adjust the cell concentration to 5x10 4 / mL, inoculate into 96-well plates at 100 μL / well; incubate the 96-well plates in a 37°C, 5% CO2 cell culture incubator for 24 hours until the cells adhere;

[0090] Drug addition: Aspirate the old culture medium, wash once with PBS, and then add 100 μL / well of the pre-prepared drug solution of each concentration to a 96-well plate. Set up 3 replicates for each concentration. Add an equal volume of complete culture medium containing 1‰ DMSO to the control group and continue culturing in a cell culture incubator for 24 hours.

[0091] Add CCK-8 reagent: Aspirate the old culture medium, wash once with PBS, and in a dark environment, add 100 μL of basal culture medium plus 10 μL of CCK-8 reagent to each well. Cover with aluminum foil and continue incubation in the incubator for another 2 hours. Measure the OD value at 450 nm using a microplate reader.

[0092] Experimental data processing

[0093] The cell growth inhibition rate was calculated according to the following formula:

[0094] Inhibition rate = 1-[(experimental group-blank group) / (control group-blank group)]*100%

[0095] According to the inhibition rate of the drug on cell proliferation at different concentrations, the IC of the compound was fitted using GraphPad Prism 8 software. 50 curve and derive IC 50 value.

[0096] Table 2 Inhibitory effect of compounds on osimertinib-resistant NSCLC cells

[0097]

[0098] As shown in Table 2, compound 20 exhibited the most significant inhibitory activity, with an IC50 value of 2.47 μM, significantly superior to the other compounds. Compounds 17 (8.83 μM) and 16 (11.53 μM) also showed moderate activity, while the IC50 values ​​of the remaining compounds (such as 13, 18, etc.) were mostly distributed in the range of 13-15 μM. Compound 20 was superior to compound 5b (2.83±0.24 μM) designed by Kshipra S. Karnik et al. ( Bioorganic Chemistry 107 (2021) 104612 ); The structural formula of compound 5b is as follows:

[0099] .

[0100] Example 16

[0101] We compared the quinoline derivative disclosed in Example 17 of the prior art (publication number CN1446212A, publication date October 1, 2003), and the molecular docking binding between Example 17 and Aurora-B (e.g. Figure 1 As shown in the figure, it can be seen that this compound has only two hydrogen bonds with Aurora-B, which is less than the number of hydrogen bonds with anlotinib and compounds 14 and 16 of the present invention. In addition, our in vitro enzyme activity experiments show that when the carbon chain length of the quinoline side chain at the seventh position of the compound is 3, it will affect the stability of the hydrogen bond with ARG81, resulting in a decrease in the inhibitory activity of the small molecule against Aurora-B.

Claims

1. A quinoline compound, characterized in that Any of the following structural formulas: 、 、 、 、 、 、 、 。 2. A pharmaceutically acceptable salt of the quinoline compound according to claim 1.

3. A pharmaceutical composition, characterized in that: The invention comprises the compound according to claim 1 and pharmaceutically acceptable excipients.

4. A pharmaceutical composition, characterized in that: Comprising the pharmaceutically acceptable salt according to claim 2 and pharmaceutically acceptable excipients.

5. A pharmaceutical composition according to claim 3 or 4, characterized in that: The dosage form of the drug is an aqueous dispersion, liquid, gel, syrup, slurry, suspension, aerosol, controlled release agent, fast dissolution agent, effervescent agent, lyophilized agent, tablet, powder, pill, dragee, capsule, delayed release agent, extended release agent, pulse controlled release agent, multiparticulate or immediate release agent.

6. Use of the compound according to claim 1 in the preparation of a medicament having dual inhibitory effects on Aurora kinase B and epidermal growth factor receptor.

7. Use of the pharmaceutically acceptable salt according to claim 2 in the preparation of a medicament having dual inhibitory effects on Aurora kinase B and epidermal growth factor receptor.

8. Use of the pharmaceutically acceptable compound of claim 1 in the preparation of a medicament for treating tumors associated with overexpression of Aurora kinase B and epidermal growth factor receptor.

9. Use of the pharmaceutically acceptable salt according to claim 2 in the preparation of a medicament for treating tumors caused by overexpression of Aurora kinase B and epidermal growth factor receptor.

10. Use of the compound according to claim 1 in the preparation of a medicament for treating non-small cell lung cancer.

11. Use of the pharmaceutically acceptable salt according to claim 2 in the preparation of a medicament for treating non-small cell lung cancer.

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