A class of compounds containing 2-aryl-4-substituted quinoline structure and preparation method and application thereof

Abstract

The present invention discloses a class of compounds containing a 2-aryl-4-substituted quinoline structure, whose structure is shown in general formula I: The class of compounds containing a 2-aryl-4-substituted quinoline structure provided by the present invention has a relatively obvious proliferation inhibition effect on lung cancer A549, colon cancer HCT116, and breast cancer MDA-MB-231. The anti-tumor activity of some compounds is significantly better than that of cisplatin, and they can be used as candidate anti-tumor drugs for more in-depth research.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a method for preparing a compound containing a 2-aryl-4-substituted quinoline structure and an application thereof in preparing an anti-tumor drug. Background Art

[0002] Malignant tumors seriously endanger human health. In recent years, the incidence and mortality of cancer in my country have increased year by year. According to the WHO Global Cancer Report: In 2018, the number of cancer cases in China was 4.056 million (accounting for about 22% of the total number of cancer cases in the world), of which 2.618 million died of cancer (accounting for about 27% of the total number of cancer deaths in the world). At present, more than 150 kinds of anti-tumor drugs are commonly used in clinical practice, but most of them have problems such as limited efficacy, large toxic side effects and serious drug resistance. Therefore, the research and development of innovative anti-tumor drugs with new structural types, new mechanisms of action, high efficiency and low toxicity has always been a hot spot in global new drug research and development.

[0003] Microtubules are important components of eukaryotic cells, and together with other proteins they assemble into a variety of structures, including spindles, centrioles, flagella, and neural tubes. Under normal physiological conditions, the polymerization and depolymerization of microtubules maintain a dynamic balance, so cell division is highly controllable and progresses in an orderly manner. This unstable dynamic characteristic makes microtubules have important regulatory functions in key cellular biological processes such as maintaining cell morphology, cell migration, cell mitosis, the transport of intracellular substances, and signal transduction. Microtubules polymerize into spindles in the early stages of cell division, and the spindles pull chromosomes toward the two poles during cell mitosis and enter the two daughter cells, thereby completing cell proliferation. The wall of the microtubule is surrounded by 14 parallel fibrils, and each fibril is formed by αβ tubulin dimers connected head to tail in sequence. This structure allows microtubules to form polar α and β ends. Due to the important role of microtubules in cell mitosis, tubulin has gradually become one of the important targets for medical workers to research and develop anticancer drugs, and tubulin inhibitors targeting tubulin have also become clinically proven effective anti-tumor drugs. The mechanism of action of this type of inhibitor is to target the microtubules in vascular endothelial cells, affect the assembly of microtubules and interfere with the normal structure and function of microtubules, prevent the formation of spindle fibers or make the spindle fibers lose power, and the sister chromatids lose the traction force for separation during mitosis, and cannot be correctly oriented in both directions, thereby interfering with the mitotic process of cells, interrupting cell mitosis and stagnating in the M phase, thereby leading to apoptosis of tumor cells, destroying the endothelial cell skeleton structure, causing selective occlusion of the tumor vascular system and large-scale damage and necrosis, and exerting an anti-tumor effect.

[0004] Studies have found that there are 8 inhibitor binding sites on tubulin. Among them, the paclitaxel site, vinblastine site and colchicine site were discovered earlier, and their mechanisms of action are relatively mature. In addition, paclitaxel and vinblastine site tubulin inhibitors have occupied an important position in the clinical treatment of tumors. Paclitaxel has become an important first-line treatment for lung cancer, breast cancer and ovarian cancer. Currently, the widely used tubulin inhibitors in clinical practice include paclitaxel, docetaxel, vinblastine and vinorelbine. However, these drugs have problems such as difficult to tolerate side effects and easy drug resistance after use, which limits the clinical use of tubulin inhibitors. However, compared with inhibitors at the paclitaxel site and vincristine site, the active cavity volume of the colchicine binding site is smaller, which is convenient for research; and the inhibitors at the colchicine site are still effective against tumor strains resistant to paclitaxel and vinblastine, and are not easy to develop drug resistance. Therefore, the development of new tubulin inhibitors with stronger activity, lower toxicity, and more effective against multidrug-resistant tumor strains has great application prospects.

[0005] In the previous work, our team used the virtual screening method to discover a class of 2-aryl-substituted quinoline derivatives as inhibitors of colchicine binding sites, and showed moderate and broad-spectrum anti-tumor activity (Bioorg Chem, 2022, 118, 105486). On this basis, this patent has structurally modified the lead structure discovered in the early stage and discovered a new class of 2-aryl-4-substituted quinoline compounds with better anti-tumor activity. Enzyme experiments have confirmed that this class of compounds can inhibit the polymerization of microtubules and have the potential to be developed into new anti-tumor drugs. Summary of the invention

[0006] The purpose of the present invention is to provide a class of compounds containing 2-aryl-4-substituted quinoline structures, which have a new skeleton structure and excellent anti-tumor activity and can be used to prepare anti-tumor drugs.

[0007] The second object of the present invention is to provide an application of the compound containing a 2-aryl-4-substituted quinoline structure in the preparation of anti-tumor drugs.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0009] The first aspect of the present invention provides a class of compounds containing a 2-aryl-4-substituted quinoline structure, the structure of which is shown in general formula I:

[0010]

[0011] In the general structural formula, Y is -OH, -Cl, -Br, -N3, -NH2; R1 and R2 are -Me, -OMe, -F, -OCF3 substituents; the 2-aryl group of quinoline can be a benzene ring or a heterocycle such as pyrrole, furan or benzofuran.

[0012] In the structural formula, n=1 represents a connected carbon atom, and n=0 represents a disconnected structure.

[0013] Most preferably, the structure of the general formula I or its pharmaceutically acceptable salt is selected from one of the following structures:

[0014]

[0015] The second aspect of the present invention provides a use of the compound having a 2-aryl-4-substituted quinoline structure or a pharmaceutically acceptable salt thereof in the preparation of an anti-tumor drug.

[0016] The 2-aryl-4-substituted quinoline structure compound is preferably compounds 1 to 14, among which compounds 4 and 6 are most preferred.

[0017] The tumor is lung cancer, colon cancer or breast cancer.

[0018] Due to the adoption of the above technical solution, the present invention has the following advantages and beneficial effects:

[0019] The 2-aryl-4-substituted quinoline structure compounds provided by the present invention have a relatively obvious proliferation inhibition effect on breast cancer MDA-MB-231, lung cancer A549 and intestinal cancer HCT116, and the anti-tumor activity of some compounds is stronger than that of the clinical drug cisplatin. For example, compound 4 has the best overall anti-tumor activity, with a half-inhibitory concentration IC of 1.37 for lung cancer, breast cancer and intestinal cancer. 50 The results showed that the concentrations of cisplatin in the serum of 1 μg / mL and 1 μg / mL were 0.86 μM, 0.65 μM and 0.90 μM, respectively, which were better than those of the clinical drug cisplatin (IC 50 >5μM), and can be used as a lead structure for anti-tumor research. The compounds provided by the present invention have a new skeleton structure, containing hydroxyl groups, amino groups, etc., which are conducive to further rapid derivatization; compared with previous research results, these compounds have a new skeleton structure; and this type of compound has a strong anti-tumor activity and can be used for the development of anti-tumor drugs.

[0020] The compound containing a 2-aryl-4-substituted quinoline structure provided by the invention has a simple synthesis route, and the synthetic raw materials are easily available and the synthetic method is easy to implement.

[0021] The use of the compound containing a 2-aryl-4-substituted quinoline structure provided by the present invention in the preparation of an anticancer drug preparation can provide more options for clinical treatment.

[0022] The compounds with 2-aryl-4-substituted quinoline structure provided by the present invention have a simpler chemical synthesis route, and some of the compounds show better in vitro anti-tumor activity than the clinical drug cisplatin. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 A is the experiment of compound 4 inhibiting tubulin polymerization.

[0024] Figure 1 B is the experiment of colchicine inhibiting tubulin polymerization.

[0025] Figure 1 C is the molecular docking diagram of compound 4 to the colchicine binding site of tubulin.

[0026] Figure 1 D is the superposition image of compound 4 and colchicine at the binding site. DETAILED DESCRIPTION

[0027] In order to explain the present invention more clearly, the present invention is further described below in conjunction with preferred embodiments. It should be understood by those skilled in the art that the content described below is illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention.

[0028] The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturers.

[0029] The chemical structural formulas of the compounds prepared in the following examples are: 1 H-NMR, 13 The C-NMR and HRMS data are shown in Table 1, wherein numbers 1 to 14 correspond to compounds 1 to 14 prepared in Examples 1 to 14, respectively.

[0030] Table 1. Chemical structures of target compounds 1 to 10. 1 H-NMR, 13 C-NMR and HRMS data

[0031]

[0032]

[0033]

[0034]

[0035] Example 1

[0036] Synthesis of compound 1

[0037]

[0038] Step 1: Take 5-methoxyisatin (150 mg, 0.85 mmol) and 2,3-dihydro-1H-inden-1-one (112 mg, 0.85 mmol), KOH (143 mg, 2.55 mmol), add 15 ml EtOH, heat under reflux at 80 ° C and react overnight. Add dilute hydrochloric acid to adjust the pH to 2. Filter to obtain a white solid (189 mg, yield 57%).

[0039]

[0040] Take the compound obtained in step 1 (37 mg, 0.127 mmol) and NaBH4 (14.2 mg, 0.375 mmol) and add THF (3 mL). Lower the temperature to 0 °C and stir. Add BF3·OEt2 (60 μL) dropwise within 1 h. Heat to room temperature and react for 16 h. Add 2 ml of 1% sodium hydroxide aqueous solution. Stir for 3 h. Evaporate the solvent under reduced pressure and purify the residue by silica gel column chromatography (DCM: MeOH = 50: 1). Obtain compound 1 as a brown solid (11 mg, yield 31%)

[0041] Example 2

[0042] Synthesis of compound 2

[0043]

[0044] Referring to Example 1, the reaction gave compound 2 as a brown solid (12 mg, yield: 29%).

[0045] Example 3

[0046] Synthesis of compound 3:

[0047]

[0048] Step 1: Take 5-methoxyisatin (150 mg, 0.85 mmol), 1-(benzo[d][1,3]dioxol-5-yl)ethan-1-one (140 mg, 0.85 mmol), KOH (143 mg, 2.55 mmol), add 15 ml of EtOH and heat under reflux at 80°C to react overnight.

[0049] Dilute hydrochloric acid was added to adjust the pH to 2. A yellow solid (186 mg, yield = 68%) was obtained by suction filtration.

[0050]

[0051] Step 2: Take the compound obtained in Step 1 (40 mg, 0.12 mmol) in THF (3 mL), stir at 0°C and add LiAlH4 (0.24 mmol), heat to room temperature and react for 30 min. Add anhydrous methanol to quench, evaporate the solvent under reduced pressure, and purify the residue by silica gel column chromatography (DCM: MeOH = 30: 1). Compound 3 is obtained as a yellow solid (27 mg, yield 73%)

[0052] Example 4

[0053] Synthesis of compound 4

[0054]

[0055] Referring to Example 3, compound 4 was obtained as a pale white solid (27 mg, yield: 66%).

[0056] Example 5

[0057] Synthesis of compound 5

[0058]

[0059] Referring to Example 3, compound 5 was obtained as a white solid (21 mg, yield was 51%).

[0060] Example 6

[0061] Synthesis of compound 6

[0062]

[0063] Referring to Example 3, compound 6 was obtained as a white solid (20 mg, yield was 40%).

[0064] Example 7

[0065] Synthesis of compound 7

[0066]

[0067] Referring to Example 3, compound 7 was obtained as a white solid (25 mg, yield was 59%).

[0068] Example 8

[0069] Synthesis of compound 8

[0070]

[0071] Referring to Example 3, compound 8 was obtained as a pale white solid (25 mg, yield: 81%).

[0072] Example 9

[0073] Synthesis of compound 9

[0074]

[0075] Referring to Example 3, compound 9 was obtained as a pale white solid (24 mg, yield was 83%).

[0076] Example 10

[0077] Synthesis of compound 10

[0078]

[0079] Referring to Example 3, compound 10 was obtained as a pale white solid (18 mg, yield was 51%).

[0080] Embodiment 11

[0081] Synthesis of compound 11

[0082]

[0083] Compound 4 (20 mg, 0.059 mmol) was added to 5 ml of DCM, stirred at 0°C and SOCl2 (172 μL) was added, and the reaction was continued for 3 h. The solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM: MeOH = 25: 1). Compound 11 was obtained as a yellow solid (11 mg, yield 52%).

[0084] Example 12

[0085] Synthesis of compound 12

[0086]

[0087] Compound 4 (40 mg, 0.12 mmol) and NBS (43 mg, 0.24 mmol) were added to THF (3 mL), stirred at 0°C and PPh3 (62 mg, 0.24 mmol) was added, heated to room temperature, and reacted for 12 h. The solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM: MeOH = 25: 1). Compound 12 was obtained as a yellow solid (26 mg, yield 54%).

[0088] Embodiment 13

[0089] Synthesis of compound 13

[0090]

[0091] Compound 12 (20 mg, 0.05 mmol) and NaN3 (6.5 mg, 0.1 mmol) were added to 1 ml of DMF and stirred at room temperature for 12 h. The solvent was evaporated under reduced pressure and the residue was purified by silica gel column chromatography (DCM: MeOH = 30: 1) to obtain compound 13 as a brown solid (11 mg, yield 60%).

[0092] Embodiment 14

[0093] Synthesis of compound 14

[0094]

[0095] Compound 13 (30 mg, 0.08 mmol), PPh3 (44 mg, 0.17 mmol) and H2O (200 μL) were added to 3 ml THF and stirred at room temperature for 12 h. The solvent was evaporated under reduced pressure and the residue was purified by silica gel column chromatography (DCM: MeOH = 30: 1). Compound 14 was obtained as a brown solid (12 mg, yield 44%).

[0096] Embodiment 15

[0097] Antitumor activity test of the compounds of the present invention.

[0098] The compounds of the present invention were subjected to a tumor cell proliferation inhibition test using conventional CKK-8 method.

[0099] The cell lines used were lung cancer A549, colon cancer HCT116, and breast cancer MDA-MB-231, all purchased from the cell bank of Shanghai Institutes for Biological Sciences. The culture medium was DMEM+10% NBS+double antibody.

[0100] Sample solution preparation: After the test compound is dissolved in DMSO (Merck), a stock solution with a concentration of 10 mM is prepared. The stock solution is diluted with culture medium to prepare the final concentrations of the drug at 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM, 1.56 μM, 0.78 μM, and 0.39 μM.

[0101] The concentration of 8 × 10 4 100 μL of cell suspension with a density of 100 cells / mL, i.e. 8000 cells / well, was placed in a 37°C, 5% CO2 incubator. After 24 hours, the upper culture medium was removed, and the culture medium containing the sample and the reference solution were added, 100 μL / well, and incubated at 37°C for 72 hours. 10 μL of CKK-8 was added to each well and placed in an incubator. After 1 hour, the OD value at 570 nm was measured using a MK-2 fully automatic microplate reader to calculate the half inhibitory concentration IC 50 .

[0102] The anti-tumor activities of some preferred compounds are shown in Table 2, where samples 1 to 14 refer to the compounds prepared in the corresponding examples, such as compound 1 refers to the compound obtained in Example 1, and so on. Positive drugs are colchicine and cisplatin.

[0103] Table 2. Half inhibitory concentration IC of some compounds of the present invention on tumor cells 50 (Unit: μM)

[0104]

[0105]

[0106] The results in Table 2 show that most of the compounds of the present application exhibit good anti-tumor activity, and have excellent proliferation inhibition effects on lung cancer A549, colon cancer HCT116 and breast cancer MDA-MB-231. The anti-tumor activity of some compounds is better than that of cisplatin, such as the half-inhibitory concentration IC of compounds 4, 6, and 7 on lung cancer A549, colon cancer HCT116 and breast cancer MDA-MB-231. 50 All of them were lower than 5 μM, showing a broad-spectrum and excellent anti-tumor activity, especially compound 4, whose half inhibitory concentration IC 50 The half inhibitory concentrations of compound 3 and compound 10 on the above three tumor cells were all lower than 1 μM. 50 All of them are lower than 12 μM, producing relatively excellent anti-tumor activity. Combined with chemical structure analysis, the above five compounds have the following common features: 1. Y is -OH; 2. n is 0; 3. The 2-aryl group of quinoline contains a benzene ring (three methoxy groups at the 3, 4, and 5 positions of the benzene ring have the best effect). Therefore, the compound skeleton with the above structural characteristics is the advantageous structural skeleton for the compound containing the 2-aryl-4-substituted quinoline structure provided by the present invention to produce excellent anti-tumor activity, and can be used for the development of anti-tumor drugs.

[0107] Example 16

[0108] Tubulin polymerization inhibition assay

[0109] This experiment was based on our published paper (Bioorg Chem, 2022, 118, 105486). Tubulin isolated from pig brain was dissolved in PEM buffer containing 1 mM GTP, 5% glycerol and the compound. Colchicine was used as a positive control. The absorbance of the mixture at 340 nm was detected at 37 ° C using a SPECTRAMAX 190 spectrophotometer. The calculation was performed using the stable absorbance value. The experiment was performed twice, and the IC was calculated accordingly. 50 value.

[0110] As attached Figure 1 As shown: Compound 4 can inhibit the polymerization of tubulin ( Figure 1 A), IC 50 The IC value of the positive drug colchicine is 13.5 μM. 50 8.1μM( Figure 1 B). Molecular docking results showed that compound 4 could be well inserted into the colchicine binding site of tubulin, and its hydroxyl group formed a hydrogen bond interaction with amino acid Asn 101, which enhanced the binding ( Figure 1 C).

Claims

1. A compound containing a 2-aryl-4-substituted quinoline structure, characterized in that: The structure is shown in general formula I: The compound containing a 2-aryl-4-substituted quinoline structure is one of the following structures:

2. A method for preparing a compound containing a 2-aryl-4-substituted quinoline structure as claimed in claim 1, characterized in that: The following steps are involved: Compounds II and III in a molar ratio of 1:1 and 3 equiv of KOH were added to EtOH, and the mixture was heated under reflux at 80°C for overnight reaction; after the reaction, dilute hydrochloric acid was added to adjust the pH to 2, and compound IV was obtained by suction filtration; 1 equiv of compound IV and 3 equiv of NaBH4 were added to THF, the temperature was lowered to 0°C, 3.8 equiv of BF3·OEt2 was added dropwise within 1 hour and stirred, and then the mixture was heated to room temperature for 16 hours; after the reaction was complete, 4 equiv of 1% aqueous sodium hydroxide solution was added and stirred for 3 hours; the solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography, wherein DCM:MeOH=50:1, to obtain the target compound V; compound V is the target skeleton I with n=1; Compounds II and VI in a molar ratio of 1:1 and 3 equiv of KOH were added to EtOH, and the mixture was heated to reflux at 80°C for overnight reaction; after the reaction, dilute hydrochloric acid was added to adjust the pH to 2, and the compound VII was obtained by suction filtration; compound VII was added to THF, stirred at 0°C, and 2 equiv of LiAlH4 was added, and the mixture was heated to room temperature for 30 min; anhydrous methanol was added to quench the mixture, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography, wherein DCM:MeOH=30:1, to obtain compound VIII; compound VIII is the target skeleton I with n=0; Compound VIII with a molar ratio of 1:2 is reacted with SOCl2 in dichloromethane under ice bath to obtain compound IX in which Y is substituted with chlorine; Compound VIII, NBS and 2 equiv of PPh3 with a molar ratio of 1:2:2 are added to dichloromethane and reacted at room temperature to obtain target compound IX in which Y is substituted with bromine; target compound IX in which Y is substituted with bromine is reacted with sodium azide in DMSO solvent at room temperature to obtain compound X; the azide group of compound X is further reacted with triphenylphosphine in a mixed solvent of tetrahydrofuran and water to obtain target compound XI; wherein compound IX, X and XI are target skeleton I in which n=0 and Y is chlorine or bromine, azide or amino respectively; Another compound of formula III and formula VI that reacts with compound II is 3. Use of a compound containing a 2-aryl-4-substituted quinoline structure according to claim 1 in the preparation of anti-tumor drugs, characterized in that: The tumor is lung cancer, breast cancer, or colon cancer.

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