Use of quinoline alkaloid derivatives for the preparation of medicaments for the treatment of cancer

Abstract

The application belongs to the field of medicine, and particularly relates to application of a series of quinoline base derivatives in preparation of anti-tumor drugs. It is found through in-vitro MTT anti-tumor activity research that the quinoline base derivatives have good inhibitory effect on human breast cancer, human cervical cancer, gastric cancer, colorectal cancer, liver cancer and the like, and the treatment effect is better than that of 5-fluorouracil, a commonly used tumor chemotherapy drug. Therefore, the quinoline base derivatives can be used for preparing anti-tumor drugs, and have good application prospect.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to the application of a series of quinoline base derivatives in the preparation of antitumor drugs. Background Technology

[0002] Cancer is a disease caused by the excessive proliferation of cells due to a loss of normal regulation. To date, cancer remains the second leading cause of death worldwide and a major public health problem. Current treatments for cancer include surgery, chemotherapy, radiotherapy, and targeted drug therapy, but existing treatments and early detection methods have not effectively controlled its incidence and high mortality rate.

[0003] Alkaloids are important natural products and, as a class of natural traditional Chinese medicine components, they exhibit significant anti-cancer effects through multiple mechanisms, including inhibiting cell proliferation, metastasis, and angiogenesis; altering cell morphology; promoting apoptosis and autophagy; triggering cell cycle arrest; and regulating various cancer-related genes and pathways. However, current chemotherapy drugs, including pentafluorouracil and topotecan, have not achieved the expected efficacy and still suffer from serious side effects and drug resistance, which limits the development of chemotherapy drugs. For the development of drugs for metastatic diseases that are difficult to cure with chemotherapy, including novel cytotoxic agents, remains a potentially effective method for cancer treatment. Therefore, the synthesis and development of highly effective and low-toxicity chemotherapy drugs is a necessary cancer treatment strategy.

[0004] Due to their rational design and advanced synthesis, quinoline derivatives have found wide application in medicinal chemistry, particularly in the development of anticancer drugs or drug candidates. This invention discovers a class of quinoline alkaloid derivatives that can effectively inhibit the proliferation of breast cancer, cervical cancer, liver cancer, colorectal cancer, and gastric cancer cells, with effects superior to commonly used clinical drugs like 5-fluorouracil. They also exhibit low toxicity to normal intestinal epithelial cells and good safety profile. Therefore, these quinoline alkaloid derivatives have promising applications as chemotherapeutic agents for cancer treatment. Summary of the Invention

[0005] In light of the above background, this invention has discovered a class of quinoline base derivatives that can effectively inhibit the proliferation of common cancer cells, and whose efficacy is superior to that of the commonly used clinical drug 5-fluorouracil, exhibiting low toxicity to normal intestinal epithelial cells and good safety. Specifically, it includes the following:

[0006] In a first aspect, the present invention provides the use of a quinoline alkaloid derivative or a pharmaceutically acceptable salt thereof in the preparation of an antitumor drug, wherein the structural formula of the quinoline alkaloid derivative is shown in formula (I) or formula (II) below:

[0007]

[0008] R1 is selected from hydrogen, -CF3, R2 is selected from hydrogen,

[0009]

[0010] R3 is selected from hydrogen, -CF3;

[0011] R4-R6 are selected from hydrogen, methyl, and halogen, respectively;

[0012] R 1 -R 3 They are selected from hydrogen, halogen, methoxy, and methyl, respectively;

[0013] X is selected from Mn, Mg, or Ni;

[0014] n is 0 or 1.

[0015] Preferably, the structural formula of the quinoline base derivative includes:

[0016]

[0017] Preferably, the tumor includes breast cancer, cervical cancer, gastric cancer, colorectal cancer, and liver cancer. Preferably, the structural formula of the quinoline base derivative includes:

[0018]

[0019] Preferably, the tumor is colon cancer.

[0020] Preferably, the structural formula of the quinoline base derivative includes:

[0021] Preferably, a quinoline derivative or a pharmaceutically acceptable salt thereof is added to a pharmaceutically acceptable carrier and / or excipients to form any pharmaceutically acceptable dosage form.

[0022] Preferably, the dosage form includes any one of tablets, sprays, granules, capsules, oral liquids, injections, and suspensions.

[0023] The beneficial effects of this invention are as follows: In vitro MTT assays revealed that the quinoline derivatives described in this application exhibit good inhibitory effects against human breast cancer, cervical cancer, gastric cancer, colorectal cancer, and liver cancer, with therapeutic effects superior to commonly used clinical chemotherapy drugs such as 5-fluorouracil. Therefore, the quinoline derivatives described in this application can be used to prepare antitumor drugs and have promising application prospects. Attached Figure Description

[0024] Figure 1Effects of quinoline derivatives 10 and 12 on HCT116 and Caco-2 cell clones;

[0025] Figure 2 Effects of quinoline derivatives 10 and 12 on apoptosis in HCT116 and Caco-2 cells. Detailed Implementation

[0026] To make the objectives, technical solutions, and advantages of this invention clearer, the various embodiments of this invention will be described in detail below with reference to the accompanying drawings. However, those skilled in the art will understand that many technical details have been provided in the embodiments of this invention to facilitate a better understanding of this application. However, the technical solutions claimed in this application can be implemented even without these technical details and various variations and modifications based on the following embodiments.

[0027] The human cancer cell lines used in the following examples include colorectal cancer cell line HCT116, gastric cancer cell line AGS, liver cancer cell line SMMC-7721, human breast cancer cell line MDA-MB-231, human cervical cancer cell line HeLa, and human normal intestinal epithelial cell line HIEC; HCT116 and AGS were purchased from ATCC (USA), and SMMC-7721, MDA-MB-231, HeLa, and HIEC were from the School of Pharmacy, Lanzhou University. HCT116, AGS, SMMC-7721, HeLa, and HIEC cell lines were cultured in 1640 medium (1640, Solarbio Invitrogen Corp., Beijing, China); the MDA-MB-231 cell line was cultured in high glucose medium (DMEM, Solarbio Invitrogen Corp., Beijing, China); the culture conditions were 37°C, 5% CO2, and all cells were supplemented with 10% fetal bovine serum (FBS), 100 U / mL penicillin, and 100 mg / mL streptomycin.

[0028] The numbering and structural formulas of the quinoline base derivatives described in the following examples are shown in Table 1 below:

[0029] Table 1. Numbering and structural formulas of quinoline base derivatives

[0030]

[0031]

[0032] The synthetic methods of the selected compounds 10, 11, 12, 13, 17, and 20 are referenced in the following reference: Yang, YD, He, YH, Ma, KY, Li, H., Zhang, ZJ, Sun, Y., ... Liu, YQ (2021). Design and Discovery of Novel Antifungal Quinoline Derivatives with Acylhydrazide as a Promising Pharmacophore. J Agric Food Chem, 69(30), 8347-8357.

[0033] The synthetic methods of the selected compounds 198, 200, 207, and 208 are referenced in: Yin, XD, Ma, KY, Wang, YL, Sun, Y., Shang, XF, Zhao, ZM, ... Liu, YQ (2020). Design, Synthesis, and Antifungal Evaluation of 8-Hydroxyquinoline Metal Complexes against Phytopathogenic Fungi. J Agric Food Chem, 68(40), 11096-11104.

[0034] The synthetic methods for selected compounds 273, 323, 324, and 329 are referenced in: Chen, YJ, Ma, KY, Du, SS, Zhang, ZJ, Wu, TL, Sun, Y.,...Tang, C. (2021). Antifungal Exploration of Quinoline Derivatives against Phytopathogenic Fungi Inspired by Quinine Alkaloids. J Agric Food Chem, 69(41), 12156-12170.

[0035] Example 1: In vitro anticancer activity of quinoline alkaloid derivatives against different human cancer cell lines

[0036] The in vitro anticancer activity of the 14 quinoline derivatives listed in Table 1 against different human cancer cell lines (including AGS, HCT116, SMMC-7721, MDA-MB-231, and HeLa) was studied using the MTT assay, with 5-fluorouracil (5-FU), a commonly used clinical chemotherapy drug, as a positive control. The specific method was as follows:

[0037] Different cancer cells were collected in 96-well plates (1×10⁻⁶). 4 Cells were cultured in 96-well plates (100 μL / cell) and pre-incubated at 37°C for 12 h to allow them to adhere. Cells were then treated with different doses of quinoline alkaloid derivatives for 48 h. 10 μL of 5 mg / mL MTT solution was added to each well, and the plates were incubated for another 4 h in a cell culture incubator. After incubation, the liquid in the 96-well plates was aspirated, and 100 μL of DMSO solution was added to each well. The plates were then incubated on a shaker at 120 rpm for 30 min. The absorbance at 490 nm was measured using a microplate reader, and the IC50 values ​​of different quinoline alkaloid derivatives against cancer cells were calculated. 50 .

[0038] Quinoline alkaloid derivatives and 5-FU positive controls showed 48-hour IC50 values ​​for five different cancer cells. 50 The (μM) values ​​are shown in Table 2. The results indicate that: ① Among all experimental cancer cell types, colorectal cancer cells and gastric cancer cells showed higher sensitivity to most of the quinoline base derivatives described in this application, especially in colorectal cancer cells, where the IC50 values ​​for most quinoline base derivatives were significantly higher. 50 ① The concentration reaches below 5 μM; ② Compared with 5-FU positive drugs, the quinoline derivatives described in this application have better inhibitory activity against cancer cells, and the activity is much greater than that of 5-FU; ③ The quinoline derivatives No. 10, No. 200, and No. 329 described in this application all have good inhibitory activity against cancer cells, among which, the IC50 against colorectal cancer cells is significantly higher. 50 The concentrations were 1.560μM, 2.285μM, and 2.568μM, respectively, and their anticancer activity was significantly better than that of 5-FU, about ten times that of 5-FU.

[0039] Table 2. IC50 of quinoline derivatives and 5-FU on HCT116, AGS, and HeLa tumor cells. 50 (μM)

[0040] Compound HCT116 AGS Hela SMMC7721 MDA-MB-231 HIEC 10 1.560±0.31 2.931±0.15 2.578±0.86 1.397±0.75 1.427±0.24 9.913±1.24 11 5.209±0.94 6.902±1.24 7.743±1.18 6.375±1.24 17.275±2.61 12.908±2.13 12 4.363±0.64 5.982±1.29 8.980±2.48 6.667±1.85 5.397±0.92 9.522±1.35 13 5.023±0.82 5.447±0.42 6.796±0.74 6.338±0.67 8.643±1.46 21.23±2.63 17 3.404±0.31 6.338±1.53 10.130±1.27 5.312±0.35 7.348±1.55 24.291±1.98 20 12.145±1.39 13.686±1.38 14.068±2.65 18.931±3.18 15.365±2.35 29.237±2.85 198 9.058±1.95 6.895±1.29 4.577±0.76 4.256±0.63 4.234±0.72 8.41±1.35 200 2.285±0.15 5.054±1.14 4.973±0.48 4.578±0.16 4.293±0.67 6.463±0.76 207 4.621±1.33 4.291±0.83 7.007±1.56 6.773±0.84 7.327±0.38 8.678±1.55 208 4.746±1.85 2.597±1.39 2.845±0.37 3.464±0.44 5.385±0.88 6.431±0.74 273 3.571±1.23 8.293±2.85 22.470±2.65 15.594±1.46 16.284±1.87 ＞20 323 7.137±1.58 8.024±1.56 12.797±1.37 13.345±2.45 13.247±1.38 ＞20 324 6.907±1.83 8.978±2.27 13.062±1.84 15.318±1.52 14.797±2.25 10.696±1.92 329 2.568±0.74 1.675±0.37 3.520±0.64 2.012±0.48 5.760±1.36 5.375±0.63 5-FU 20.591±2.84 31.637±2.93 ＞50 ＞50 ＞50 ＞50

[0041] The above results indicate that the quinoline derivatives described in this application have good inhibitory effects on gastric cancer, colorectal cancer, liver cancer, cervical cancer, and breast cancer, and their therapeutic effects are superior to those of the commonly used clinical drug 5-FU. Therefore, it is reasonable to infer that the above compounds have good inhibitory effects on various types of tumor cells. Thus, the quinoline derivatives described in this application can be used to prepare anticancer drugs and have promising application prospects.

[0042] Example 2: Inhibitory effect of quinoline derivatives No. 10 and No. 12 on colorectal cancer cells.

[0043] 1. MTT assay for the inhibitory effect of quinoline alkaloid derivatives on colorectal cancer cells.

[0044] HCT116 cells and Caco-2 cells were treated with different concentrations of quinoline derivative No. 10 and positive control drug 5-FU, respectively. 10 μL of 5 mg / mL MTT solution was added to each well of a 96-well plate and incubated in a cell culture incubator for 4 h.

[0045] After incubation, carefully aspirate the liquid from the 96-well plate, add 100 μL of DMSO solution to each well, and incubate on a shaker at 120 rpm for 30 min. Once the solution in the 96-well plate is homogeneous, measure the absorbance at 490 nm using a microplate reader and calculate the inhibition rate.

[0046] The results are shown in Table 1. The quinoline derivatives No. 10 and No. 12 of the present invention can significantly inhibit the proliferation of colorectal cancer cells, and their effects are significantly better than those of the positive control drug 5-FU.

[0047] 2. MTT assay for the toxicity of quinoline alkaloid derivatives to normal intestinal epithelial cells (HIEC cells).

[0048] The toxicity of quinoline derivatives to normal intestinal epithelial cells (HIEC cells) was detected using the method described in section 1 above.

[0049] The results are shown in Table 2. The quinoline derivatives 10 and 12 of the compounds obtained in this invention showed low toxicity to normal intestinal epithelial cells (HIEC cells), with IC50 values ​​of [missing information]. 50 Greater than 50 μM.

[0050] 3. Cell cloning experiments

[0051] (1) When the density of HCT116 cells and Caco-2 cells reaches 80%-90%, digest them according to the above steps. HCT116 cells are seeded at a density of 500 cells / well and Caco-2 cells are seeded at a density of 700 cells / well (24-well plate). The cells are then placed in a cell culture incubator for about 24 hours.

[0052] (2) Quinoline derivatives No. 10 and No. 12 were respectively used with their IC50. 50 Cells were treated with concentrations of 1 / 2, 1, and 2 times the concentration;

[0053] (3) Treat cells with the compound for 7-10 days, and change the drug-containing culture medium every 3 days;

[0054] (4) Discard the culture medium and carefully wash the 24-well plate with PBS;

[0055] (5) Add 300 μL of 4% paraformaldehyde to fix the cells and fix them at room temperature for 30 min;

[0056] (6) Remove the fixative, wash the 24-well plate with PBS, and then add 0.1% crystal violet staining solution for 20 min.

[0057] (7) Discard the staining solution, wash the 24-well plate with PBS until clear, air dry the 24-well plate, and observe the number of cell clones;

[0058] The results are as follows Figure 1 As shown, A represents the inhibition of HCT116 cell proliferation after 10 days of treatment with different concentrations of quinoline derivative No. 10; B represents the inhibition of HCT116 cell proliferation after 10 days of treatment with different concentrations of quinoline derivative No. 12; C represents the inhibition of Caco-2 cell proliferation after 10 days of treatment with different concentrations of quinoline derivative No. 10; and D represents the inhibition of Caco-2 cell proliferation after 10 days of treatment with different concentrations of quinoline derivative No. 12. The results indicate that the quinoline derivatives No. 10 and No. 12 described in this invention can significantly inhibit the clonal proliferation of colorectal cancer cells.

[0059] 4. Apoptosis experiment

[0060] (1) When the density of HCT116 cells and Caco-2 cells reaches 80%-90%, digest them according to the above steps, using 4.0×10⁻⁶ cells respectively. 5 CE11 / hole and 4.5×10 5 The ce11 cells were seeded at a density of 6 wells in a cell culture incubator and cultured for about 24 hours.

[0061] (2) Quinoline derivatives No. 10 and No. 12 were respectively used with their IC50. 50 Cells were treated with 1 / 2, 1, and 2 times the concentration for 48 hours;

[0062] (3) Transfer the culture medium in the 6-well plate to a 10 mL centrifuge tube, wash the cells twice with PBS, 1 mL each time, and collect the washing solution into a 10 mL centrifuge tube;

[0063] (4) Digest the cells with trypsin without EDTA. The digestion time is generally about 1 minute. Add the collected culture medium and mix the cells. Centrifuge at 1800 r / min for 5 minutes.

[0064] (5) Discard the supernatant, add 1 ml of PBS pre-cooled at 4℃, resuspend the cells, and collect the cells by centrifugation;

[0065] (6) Discard the supernatant, dilute the binding buffer with deionized water at a ratio of 1:9, add it to the centrifuge tube, and adjust its concentration to 1-5 × 10⁻⁵. 6 ce11 / mL;

[0066] (7) Take 100 μL of cell suspension into a 10 mL flow cytometer, add 5 μL of Annexinv / Alexa Fluor 488, mix well, and incubate at room temperature in the dark for 5 min.

[0067] (8) Add 5 μL of propidium iodide solution (PI) and 400 μL of PBS solution, and immediately perform flow cytometry detection and data analysis.

[0068] Analysis results as follows Figure 2 As shown, A represents the effect of different concentrations of quinoline derivative No. 10 on apoptosis of HCT116 cells; B represents the effect of different concentrations of quinoline derivative No. 12 on apoptosis of HCT116 cells; C represents the effect of different concentrations of quinoline derivative No. 10 on apoptosis of Caco-2 cells; and D represents the effect of different concentrations of quinoline derivative No. 12 on apoptosis of Caco-2 cells. The results indicate that quinoline derivatives No. 10 and No. 12 described in this application can significantly promote apoptosis in colorectal cancer cells in a concentration-dependent manner after treatment of HCT116 and Caco-2 cells.

[0069] The embodiments described above are merely illustrative of implementation methods of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make other improvements without departing from the concept of the present invention, and these improvements all fall within the protection scope of the present invention.

Claims

1. The use of quinoline base derivatives or pharmaceutically acceptable salts thereof in the preparation of antitumor drugs, characterized in that, The structural formula of the quinoline base derivative is shown in formula (Ⅰ):

2. The application as described in claim 1, characterized in that, The tumors include breast cancer, cervical cancer, stomach cancer, colorectal cancer, and liver cancer.

3. The application as described in claim 2, characterized in that, The tumor is colon cancer.

4. The application as described in any one of claims 1-3, characterized in that, Quinoline derivatives or their pharmaceutically acceptable salts are added to pharmaceutically acceptable excipients to form any pharmaceutically acceptable dosage form.

5. The application as described in claim 4, characterized in that, The dosage form includes any one of the following: tablets, sprays, granules, capsules, oral liquids, injections, and suspensions.

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