CDDO quinoline derivatives inhibiting hypoxia-inducible factor 1 alpha and preparation method and application thereof

Abstract

The application discloses a CDDO quinoline derivative with a structure as shown in formula III, R is selected from R1 is selected from R2 is selected from n is selected from an integer from 2 to 5. Pharmacological experimental researches show that the CDDO quinoline derivative can reduce the content of HIF-1 alpha protein of tumor cells, can inhibit tumor cell proliferation under hypoxia, and the inhibition effect on the proliferation of various tumor cells is significantly better than that of CDDO and CDDO-Me. The application discloses application of the CDDO quinoline derivative in preparation of an anti-tumor drug.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemistry, and relates to CDDO quinoline derivatives and a preparation method and application thereof, in particular to CDDO quinoline derivatives for inhibiting hypoxia-inducible factor 1 alpha and a preparation method and application thereof. BACKGROUND

[0002] Oleic acid (OA) is a widely existing pentacyclic triterpenoid natural product in plants, and has weak anti-tumor, anti-inflammatory, liver protection and other biological activities. A series of derivatives with significantly improved anti-tumor activity are obtained by researching and modifying OA, and the CDDO series is more prominent among them, including CDDO, CDDO-Me (bardoxolone methyl) and CDDO-Im.

[0003] CDDO-Me is a star molecule in the CDDO derivatives, and has strong anti-tumor activity, and has entered clinical experiments for treating pancreatic cancer, melanoma and the like several times. Therefore, designing derivatives of CDDO as a parent compound has great prospects.

[0004] Solid tumors are often in a hypoxic state (oxygen content less than <21%), and need to adapt to the hypoxic environment to maintain their rapid growth. Hypoxia-inducible factor (HIF) as a transcription factor is the main regulator of the cellular hypoxic response. Under hypoxic conditions, HIF-1 alpha can stably exist, and through the transcriptional activation of HIF-1 on the downstream effector gene, the tumor tissue can trigger and develop a series of complex basic biological mechanisms to cope with hypoxic stress and promote survival, such as angiogenesis, glycolysis, hematopoiesis, metastasis, invasion, chemotherapy resistance and radiotherapy resistance. Therefore, CDDO will be used as a lead compound to design a series of compounds to inhibit HIF-1 alpha protein, and then inhibit the development of tumors. SUMMARY

[0005] The purpose of the present application is to modify the structure by taking CDDO as a lead, and a CDDO quinoline derivative is provided. Pharmacological studies show that the anti-tumor activity of the CDDO derivative of the present application is excellent, and the content of HIF-1 alpha protein of tumor cells is reduced.

[0006] The purpose of the present application is achieved by the following technical solutions:

[0007] The CDDO quinoline derivative has the structure shown in formula III:

[0008]

[0009] wherein R is selected from R1 is selected from R2 is selected from n is an integer selected from 2 to 5.

[0010] Preferably, R is selected from R1 is selected from R2 is selected from n is an integer selected from 2 to 5; but not including: R is selected from R1 is selected from n = 5.

[0011] More preferably, R is selected from R1 is selected from R2 is selected from n is an integer selected from 2 to 5; but not including: R is selected from R1 is selected from n = 5.

[0012] As one of the technical solutions of the CDDO quinoline derivative of the present application, the structure of the CDDO quinoline derivative is shown in formula I:

[0013]

[0014] R1 is selected from n is an integer selected from 2 to 5.

[0015] Preferably, R1 is selected from n is an integer selected from 2 to 5; but not including: R is selected from R1 is selected from n = 5.

[0016] As one of the technical solutions of the CDDO quinoline derivative of the present application, the structure of the CDDO quinoline derivative is shown in formula II:

[0017]

[0018] R2 is selected from n is an integer selected from 2 to 5.

[0019] Specifically, the CDDO quinoline derivative of the present application is selected from the compounds with the following structures:

[0020]

[0021]

[0022] Another object of the present application is to provide a preparation method of the CDDO derivative,

[0023] When R is selected from , the synthesis route is as follows:

[0024]

[0025] wherein R1, n are as described above; R1-OH is selected from

[0026] comprising the following steps:

[0027] Step (1), with potassium carbonate as catalyst, N,N-dimethylformamide (DMF) as reaction solvent, quinoline hydroxyl compound represented by formula R1-OH reacts with dibromoalkane represented by formula at room temperature to obtain intermediate d; wherein the molar ratio of quinoline hydroxyl compound to dibromoalkane is 1:3-1:5; the volume ratio of dibromoalkane to potassium carbonate is 1:1-1:1.5;

[0028] Step (2), with potassium carbonate, potassium iodide as catalyst, N,N-dimethylformamide (DMF) as reaction solvent, intermediate d reacts with CDDO at room temperature to obtain CDDO quinoline derivative; wherein the molar ratio of CDDO to intermediate d is 1:1.1-1:1.5; the molar ratio of CDDO to potassium carbonate is 1:4-1:6; the molar ratio of CDDO to potassium iodide is 1:0.3-1:0.5.

[0029] when R is selected from , the synthesis route is as follows:

[0030]

[0031] wherein R1, n are as described above; selected from

[0032] comprising the following steps:

[0033] Step (1), with potassium carbonate as catalyst, N,N-dimethylformamide (DMF) as reaction solvent, quinoline hydroxyl compound represented by formula reacts with dibromoalkane represented by formula at room temperature to obtain intermediate h; wherein the molar ratio of quinoline carboxyl compound to dibromoalkane is 1:3-1:5; the volume ratio of dibromoalkane to potassium carbonate is 1:1-1:1.5;

[0034] Step (2), the intermediate d is reacted with CDDO in normal temperature to obtain CDDO quinoline derivative, with potassium carbonate and potassium iodide as catalysts and N,N-dimethylformamide (DMF) as reaction solvent; wherein, the molar ratio of CDDO to intermediate h is 1:1.1-1:1.3; the molar ratio of CDDO to potassium carbonate is 1:3-1:5; and the molar ratio of CDDO to potassium iodide is 1:0.3-1:0.5.

[0035] Another object of the present application is to provide the use of the CDDO quinoline derivative in the preparation of an anti-tumor drug.

[0036] The CDDO quinoline derivative of the present application can be used alone or in combination with clinically commonly used anti-tumor drugs such as anti-metabolic drugs, alkylating agents, anti-tumor antibiotics, anti-tumor plant drugs, hormones, and can also be used in combination with radiotherapy.

[0037] Preferably, the use is the use of the CDDO quinoline derivative in the preparation of an anti-tumor drug by inhibiting HIF-1α protein.

[0038] The tumor is melanoma, liver cancer, breast cancer, gastric cancer.

[0039] Another object of the present application is to provide the use of the CDDO quinoline derivative with the following structure in the preparation of an anti-liver cancer drug:

[0040]

[0041] Another object of the present application is to provide a pharmaceutical composition, which takes the CDDO quinoline derivative as an effective component, and is supplemented with a pharmaceutically acceptable carrier to be made into any pharmaceutically acceptable dosage form.

[0042] The CDDO quinoline derivative can be made into different dosage forms alone or in combination with one or more than one pharmaceutical carrier.

[0043] The dosage form is selected from tablets, capsules, dripping pills, granules, powders, lozenges, aqueous or oily suspensions, injections, patches, nano-preparations, for clinical oral administration, injection or topical administration. In these different preparations, the content of the CDDO quinoline derivative of the present application can be 0.1%-99.9%.

[0044] The present application has the following advantages:

[0045] The CDDO quinoline derivative of the present application has a novel structure, the preparation method has mild reaction conditions, the reagents used are low in toxicity, the raw materials are easy to obtain, the post-treatment is convenient, and the yield is high.

[0046] Pharmacological studies show that the CDDO quinoline derivative of the application can reduce the content of HIF-1α protein of tumor cells, and the inhibition effect on the proliferation of various tumor cells is significantly better than that of CDDO. Compared with CDDO and CDDO-Me, the CDDO quinoline derivative of the application has significantly enhanced inhibition effect on tumor cells under hypoxia, and the toxicity is reduced compared with CDDO. BRIEF DESCRIPTION OF DRAWINGS

[0047] Figure 1 Compound I-7 on B16-F10 cell HIF-1α protein content level immunofluorescence results.

[0048] Figure 2 Compound I-7 on B16-F10 cell HIF-1α protein content immunofluorescence column chart.

[0049] Figure 3 Compound I-7 on B16-F10 cell HIF-1α, VEGF protein content Western Bolt result chart.

[0050] Figure 4 Compound I-7 on B16-F10 cell HIF-1α protein content Western Bolt column chart.

[0051] Figure 5 Compound I-7 on B16-F10 cell VEGF protein content Western Bolt column chart. DETAILED DESCRIPTION

[0052] In order to further illustrate the present application, a series of examples are listed below. These examples are illustrative and should not be construed as limiting the present application.

[0053] Example 1

[0054] Synthesis of intermediate A1

[0055]

[0056] In a single neck flask, 3-hydroxyquinoline (2.06 mmol, 0.30 g), potassium carbonate 0.82 g (6.18 mmol), 1,2-dibromoethane 0.73 mL (8.24 mmol) were added, dissolved in 10 mL DMF, and reacted at room temperature, and the reaction was monitored by TLC. After the reaction was completed, the organic phase was rotary evaporated, and column chromatography on silica gel (eluent: petroleum ether: ethyl acetate = 10:1 v:v) was performed to obtain 3-(2-bromoethoxy)quinoline (white solid) 0.21 g, yield 40.6%.

[0057] ESI-MS: bromine isotope characteristic peaks 249.9, 251.9 [M+H] + .

[0058] Synthesis of compound 1-1

[0059]

[0060] To a solution of 0.10 g (0.22 mmol) of CDOO in 2 mL of DMF, 0.091 g of potassium carbonate (0.66 mmol) was added, followed by 0.018 g of potassium iodide (0.11 mmol), after stirring, 3-(2-bromoethoxy)quinoline 0.060 g (0.24 mmol) was added, and the reaction was carried out at room temperature until TLC detection showed that the reaction was complete, diluted with 30 mL of ethyl acetate, washed with water 3 times, the organic layer was washed with saturated sodium bicarbonate 3 times, and saturated brine 3 times, and dried over anhydrous sodium sulfate overnight. The organic phase was rotary evaporated, and column chromatography on silica gel (eluent petroleum ether: ethyl acetate = 3: 1 v:v) to obtain 0.04 g of white solid, yield 27.4%.

[0061] ESI-MS: 663.3 [M+H] + .

[0062] 1 H NMR (400 MHz, DMSO, TMS), δ ppm: δ 8.63 (s, 1H), 8.54 (d, J = 2.9 Hz, 1H), 7.98-7.93 (m, 1H), 7.91-7.86 (m, 1H), 7.83 (d, J = 2.9 Hz, 1H), 7.63-7.55 (m, 2H), 6.20 (s, 1H), 4.59-4.36 (m, 4H), 2.89 (d, J = 13.5 Hz, 1H), 2.80 (d, J = 4.7 Hz, 1H), 1.81-1.70 (m, 2H), 1.64-1.47 (m, 6H), 1.30 (s, 2H), 1.26 (s, 5H), 1.24 (s, 3H), 1.14 (s, 3H), 1.02 (s, 6H), 0.89 (s, 3H), 0.88 (s, 3H), 0.86 (s, 3H).

[0063] Example 2

[0064] Synthesis of intermediate A2

[0065]

[0066] With 1,3-dibromopropane 0.86 mL (8.24 mmol), instead of 1,2-dibromoethane in Example 1, 3-(3-bromopropoxy)quinoline (compound A2) 0.25 g, white solid, yield 45.7%.

[0067] ESI-MS: Bromine isotope characteristic peak 264.0, 266.0 [M+H] + .

[0068] Synthesis of compound I-2

[0069]

[0070] Referring to the preparation method of compound I-1 in Example 1, compound A2 was used to replace compound A1 in an equimolar amount, and other conditions were unchanged to obtain the target compound I-2 (white solid) 0.04 g, yield 26.8%.

[0071] ESI-MS: 677.3 [M+H]+.

[0072] 1 H NMR (400 MHz, DMSO, TMS), δ ppm: δ 8.60 (d, J = 2.9 Hz, 1H), 7.97-7.81 (m, 2H), 7.66 (s, 1H), 7.56 (d, J = 6.0 Hz, 2H), 7.41-7.30 (m, 1H), 6.05 (d, J = 12.8 Hz, 1H), 4.38-4.12 (m, 4H), 2.85 (d, J = 13.2 Hz, 1H), 2.69 (d, J = 4.7 Hz, 1H), 2.19 (d, J = 8.7 Hz, 2H), 1.63 (d, J = 12.0 Hz, 2H), 1.29 (d, J = 1.8 Hz, 4H), 1.23 (d, J = 12.2 Hz, 9H), 1.14 (d, J = 2.3 Hz, 3H), 1.07 (s, 3H), 0.98 (d, J = 3.0 Hz, 3H), 0.88 (s, 6H), 0.84 (d, J = 2.5 Hz, 6H).

[0073] Example 3

[0074] Synthesis of intermediate A3

[0075]

[0076] Referring to the preparation method of compound I-1 in Example 1, compound A2 was used to replace compound A1 in an equimolar amount, and other conditions were unchanged to obtain the target compound I-2 (white solid) 0.04 g, yield 26.8%.

[0077] ESI-MS: Bromine isotope characteristic peak 264.0, 266.0 [M+H] + .

[0078] Synthesis of compound I-3

[0079]

[0080] The compound I-1 was prepared according to the method described in Example 1, except that compound A3 was used instead of compound A1 in an equimolar amount. The target compound I-3 (white solid) 0.05 g was obtained in a yield of 32.8%.

[0081] ESI-MS: 691.0 [M+H] + .

[0082] 1 H NMR (400 MHz, DMSO, TMS), δ ppm: δ 8.62 (d, J = 3.0 Hz, 2H), 7.98-7.91 (m, 1H), 7.92-7.85 (m, 1H), 7.79 (d, J = 2.9 Hz, 1H), 7.62-7.52 (m, 2H), 6.22 (s, 1H), 4.18 (t, J = 6.1 Hz, 4H), 2.87 (d, J = 13.2 Hz, 1H), 2.81 (d, J = 4.8 Hz, 1H), 1.99 (s, 1H), 1.92-1.78 (m, 6H), 1.71-1.40 (m, 8H), 1.34 (s, 3H), 1.31-1.22 (m, 4H), 1.18 (d, J = 1.5 Hz, 3H), 1.15 (s, 3H), 1.03 (s, 3H), 0.92 (s, 3H), 0.87 (s, 3H), 0.85 (s, 3H).

[0083] Example 4

[0084] Synthesis of intermediate A4

[0085]

[0086] The compound A4 (white solid) 0.4 g was obtained in a yield of 66.2% by replacing 1,2-dibromoethane with 1,5-dibromopentane 1.12 mL (8.24 mmol) in Example 1.

[0087] ESI-MS: bromine isotope characteristic peaks 292.0, 294.0 [M+H] + .

[0088] Synthesis of compound I-4

[0089]

[0090] The compound I-1 was prepared according to the method described in Example 1, except that compound A4 was used instead of compound A1 in an equimolar amount. The target compound I-4 (white solid) 0.05 g was obtained in a yield of 32.2%.

[0091] ESI-MS: 705.4 [M+H] + .

[0092] 1 H NMR (400 MHz, DMSO, TMS), δ ppm: δ 8.63 (q, J = 2.9 Hz, 1H), 8.57 (s, 1H), 7.98 - 7.92 (m, 1H), 7.88 (dt, J = 7.9, 2.4 Hz, 1H), 7.79 - 7.70 (m, 1H), 7.56 (td, J = 6.4, 3.7 Hz, 2H), 6.18 (s, 1H), 4.25 - 4.06 (m, 4H), 2.87 (d, J = 13.3 Hz, 1H), 2.79 (dd, J = 11.3, 4.7 Hz, 1H), 1.86 (dt, J = 14.6, 6.6 Hz, 3H), 1.77 - 1.66 (m, 3H), 1.61 - 1.41 (m, 8H), 1.30 (d, J = 8.5 Hz, 5H), 1.27 (s, 3H), 1.24 (s, 3H), 1.18 (s, 3H), 1.11 (s, 3H), 0.98 (s, 2H), 0.91 (s, 3H), 0.90 (s, 3H), 0.87 (s, 3H).

[0093] Example 5

[0094] Synthesis of intermediate A5

[0095]

[0096] With 4-hydroxyquinoline (2.06 mmol, 0.30 g) instead of 3-hydroxyquinoline in Example 1, other conditions remain unchanged to obtain (compound A5, white solid) 0.4 g, yield 77.7%.

[0097] ESI-MS: Br isotope characteristic peaks 250.0, 252.0 [M+H] + .

[0098] Synthesis of compound I-5

[0099]

[0100] Referring to the preparation method of compound I-1 in Example 1, with equimolar amount of compound A5 instead of compound A1, other conditions remain unchanged to obtain the target compound I-5 (white solid) 0.05 g, yield 34.3%.

[0101] ESI-MS: 663.3 [M+H] + .

[0102] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 8.73 (d, J = 5.2 Hz, 1H), 8.02 (dt, J = 8.4, 1.9 Hz, 1H), 7.95 (d, J = 9.0 Hz, 1H), 7.75 (ddd, J = 8.4, 6.8, 1.5 Hz, 1H), 7.55 (ddt, J = 8.3, 6.9, 1.6 Hz, 1H), 7.08 (d, J = 5.2 Hz, 1H), 6.27 (s, 1H), 5.10 (s, 1H), 4.54 (dt, J = 10.4, 4.9 Hz, 4H), 2.91 (d, J = 13.7 Hz, 1H), 2.80 (d, J = 4.8 Hz, 1H), 1.99 (p, J = 6.9 Hz, 1H), 1.92 - 1.83 (m, 1H), 1.79 - 1.70 (m, 2H), 1.61 (d, J = 13.8 Hz, 1H), 1.45 (s, 2H), 1.25 (d, J = 10.2 Hz, 7H), 1.08 (s, 3H), 1.03 (s, 3H), 1.00 (s, 2H), 0.90 (s, 3H), 0.87 (s, 3H), 0.86 (s, 3H), 0.85 (s, 3H).

[0103] Example 6

[0104] Synthesis of intermediate A6

[0105]

[0106] With 4-hydroxyquinoline (2.06 mmol, 0.30 g) instead of 3-hydroxyquinoline in Example 2, and other conditions unchanged, compound A6 (white solid) 0.3 g, yield 54.7% was obtained.

[0107] ESI-MS: Br isotope characteristic peaks 264.0, 266.0 [M+H] + .

[0108] Synthesis of compound I-6

[0109]

[0110] With equal molar amount of compound A6 instead of compound A1 in the preparation method of compound I-1 in Reference Example 1, and other conditions unchanged, the target compound I-6 (white solid) 0.04 g, yield 26.8% was obtained.

[0111] ESI-MS: 677.3 [M+H] + .

[0112] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 8.68 (d, J = 5.2 Hz, 1H), 8.51 (s, 1H), 8.10 (dd, J = 8.5, 1.5 Hz, 1H), 7.90 (d, J = 7.9 Hz, 1H), 7.70 (ddd, J = 8.5, 6.9, 1.5 Hz, 1H), 7.52 (ddd, J = 8.2, 6.9, 1.3 Hz, 1H), 6.92 (d, J = 5.2 Hz, 1H), 6.06 (s, 1H), 4.25 (dtd, J = 15.9, 9.0, 5.2 Hz, 4H), 2.76 (d, J = 13.3 Hz, 1H), 2.61 (d, J = 4.7 Hz, 1H), 2.29 - 2.11 (m, 2H), 1.87 - 1.75 (m, 1H), 1.70 - 1.57 (m, 2H), 1.52 (d, J = 13.7 Hz, 1H), 1.46 - 1.12 (m, 10H), 1.09 (s, 3H), 1.07 (s, 3H), 0.96 (s, 3H), 0.81 (s, 4H), 0.79 (s, 3H), 0.76 (s, 3H), 0.62 (s, 3H).

[0113] Example 7

[0114] Synthesis of intermediate A7

[0115]

[0116] With 4-hydroxyquinoline (2.06 mmol, 0.30 g) instead of 3-hydroxyquinoline in Example 3, and other conditions unchanged, compound A7 (white solid) 0.4 g, yield 69.6% was obtained.

[0117] ESI-MS: Br isotope characteristic peaks 278.0, 280.0 [M+H] + .

[0118] Synthesis of compound I-7

[0119]

[0120] With equal molar amount of compound A7 instead of compound A1 in the preparation method of compound I-1 in Example 1, and other conditions unchanged, the target compound I-7 (white solid) 0.04 g, yield 26.3% was obtained.

[0121] ESI-MS: 691.4 [M+H]+.

[0122] 1H NMR (400 MHz, DMSO, TMS), δ ppm: (400 MHz, DMSO) δ 8.72 (d, J = 5.2 Hz, 1H), 8.61 (s, 1H), 8.14 (dd, J = 8.3, 1.5 Hz, 1H), 7.95 (d, J = 8.4 Hz, 1H), 7.74 (ddd, J = 8.4, 6.8, 1.5 Hz, 1H), 7.56 (ddd, J = 8.2, 6.8, 1.2 Hz, 1H), 7.02 (d, J = 5.2 Hz, 1H), 6.20 (s, 1H), 4.29 (t, J = 6.1 Hz, 2H), 4.21 (q, J = 6.1 Hz, 2H), 2.92 - 2.83 (m, 1H), 2.80 (d, J = 4.8 Hz, 1H), 1.94 (q, J = 6.5 Hz, 2H), 1.90 - 1.62 (m, 6H), 1.56 (d, J = 9.5 Hz, 5H), 1.31 (s, 4H), 1.25 (dd, J = 11.3, 5.7 Hz, 3H), 1.14 (d, J = 8.0 Hz, 9H), 1.04 (s, 3H), 0.91 (s, 3H), 0.88 (s, 3H), 0.86 (s, 3H).

[0123] Example 8

[0124] Synthesis of compound A8

[0125]

[0126] With 4-hydroxyquinoline (2.06 mmol, 0.30 g) instead of 3-hydroxyquinoline in Example 4, and other conditions unchanged, compound A8 (white solid) 0.50 g, yield 82.8% was obtained.

[0127] ESI-MS: Br isotope characteristic peaks 292.0, 294.0 [M+H] + .

[0128] Synthesis of compound I-8

[0129]

[0130] With equal molar amount of compound A8 instead of compound A1 in the preparation method of compound I-1 in Reference Example 1, and other conditions unchanged, the target compound I-8 (white solid) 0.05 g, yield 32.3% was obtained.

[0131] ESI-MS: 705.4 [M+H] + .

[0132] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 8.72 (d, J = 5.2 Hz, 1 H), 8.58 (s, 1 H), 8.14 (dd, J = 8.4, 1.5 Hz, 1 H), 7.94 (dd, J = 8.5, 1.2 Hz, 1 H), 7.73 (ddd, J = 8.5, 6.9, 1.5 Hz, 1 H), 7.56 (ddd, J = 8.3, 6.8, 1.2 Hz, 1 H), 7.00 (d, J = 5.2 Hz, 1 H), 6.18 (s, 1 H), 4.23 (td, J = 6.2, 2.4 Hz, 2 H), 4.19 - 4.05 (m, 2 H), 2.85 (dt, J = 13.7, 4.2 Hz, 1 H), 2.79 (d, J = 4.8 Hz, 1 H), 1.96 - 1.81 (m, 3 H), 1.72 (ddd, J = 17.0, 10.8, 4.2 Hz, 4 H), 1.61 (ddd, J = 17.0, 11.1, 4.2 Hz, 4 H), 1.46 (td, J = 20.3, 6.9 Hz, 5 H), 1.26 (s, 3 H), 1.24 - 1.15 (m, 3 H), 1.13 (s, 6 H), 1.08 (s, 2 H), 1.00 (s, 3 H), 0.88 (s, 3 H), 0.86 (s, 3 H), 0.84 (s, 3 H).

[0133] Example 9

[0134] Synthesis of intermediate A9

[0135]

[0136] With 6-hydroxyquinoline (2.06 mmol, 0.30 g) instead of 3-hydroxyquinoline in Example 1, other conditions remain unchanged, to obtain compound A9 (white solid) 0.42 g, yield 81.2%.

[0137] ESI-MS: Br isotope characteristic peak 250.0, 252.0 [M+H] + .

[0138] Synthesis of compound I-9

[0139]

[0140] Referring to the preparation method of compound I-1 in Example 1, with equimolar amount of compound A9 instead of compound A1, other conditions remain unchanged, to obtain the target compound I-8 (white solid) 0.05 g, yield 34.3%.

[0141] ESI-MS: 663.3 [M+H] + .

[0142] 1 H NMR (400 MHz, DMSO, TMS), δ ppm: δ 8.75 (dd, J = 4.2, 1.7 Hz, 1H), 8.62 (s, 1H), 8.25 (dd, J = 8.5, 1.7 Hz, 1H), 7.94 (d, J = 9.2 Hz, 1H), 7.49 (dd, J = 8.3, 4.2 Hz, 1H), 7.40 (d, J = 2.7 Hz, 1H), 7.30 (dd, J = 9.2, 2.8 Hz, 1H), 6.19 (s, 1H), 4.50 (q, J = 3.9 Hz, 2H), 4.37 (t, J = 4.5 Hz, 2H), 2.90 (d, J = 13.5 Hz, 1H), 2.80 (d, J = 4.7 Hz, 1H), 1.95-1.83 (m, 1H), 1.76 (td, J = 13.4, 5.3 Hz, 2H), 1.60-1.29 (m, 10H), 1.24 (s, 5H), 1.14 (s, 3H), 1.02 (s, 6H), 0.90 (s, 3H), 0.89 (s, 3H), 0.86 (s, 3H).

[0143] Example 10

[0144] Synthesis of intermediate A10

[0145]

[0146] With 6-hydroxyquinoline (2.06 mmol, 0.30 g) instead of 3-hydroxyquinoline in Example 2, and other conditions unchanged, compound A10 (white solid) 0.32 g, yield 46.4% was obtained.

[0147] ESI-MS: characteristic peaks of bromine isotope 264.0, 265.0 [M+H] + .

[0148] Synthesis of compound I-10

[0149]

[0150] With equal molar amount of compound A10 instead of compound A1 in the preparation method of compound I-1 in Example 1, and other conditions unchanged, the target compound I-10 (white solid) 0.05 g, yield 33.6% was obtained.

[0151] ESI-MS: 677.3 [M+H] + .

[0152] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 8.75 (dd, J = 4.2, 1.7 Hz, 1H), 8.52 (s, 1H), 8.26 (dd, J = 8.5, 1.7 Hz, 1H), 7.94 (d, J = 9.2 Hz, 1H), 7.50 (dd, J = 8.3, 4.2 Hz, 1H), 7.39 (dd, J = 9.1, 2.8 Hz, 1H), 7.33 (d, J = 2.8 Hz, 1H), 6.10 (s, 1H), 4.38-4.11 (m, 4H), 2.84 (dd, J = 11.3, 6.8 Hz, 1H), 2.71 (d, J = 4.7 Hz, 1H), 2.18 (q, J = 6.0 Hz, 2H), 1.94-1.84 (m, 1H), 1.77-1.58 (m, 3H), 1.51-1.22 (m, 11H), 1.15 (s, 3H), 1.08 (s, 3H), 0.98 (s, 3H), 0.88 (s, 6H), 0.85 (s, 3H), 0.84 (s, 3H).

[0153] Example 11

[0154] Synthesis of intermediate A11

[0155]

[0156] With 6-hydroxyquinoline (2.06 mmol, 0.30 g) instead of 3-hydroxyquinoline in Example 3, and other conditions unchanged, compound A11 (white solid) 0.40 g, yield 69.6% was obtained.

[0157] ESI-MS: Br isotope characteristic peaks 278.0, 280.0 [M+H] + .

[0158] Synthesis of compound I-11

[0159]

[0160] With mmol amount of compound A11 instead of compound A1 in the preparation method of compound I-1 in Example 1, and other conditions unchanged, the target compound I-11 (white solid) 0.05 g, yield 32.9% was obtained.

[0161] ESI-MS: 691.4 [M+H] + .

[0162] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 8.73 (dd, J = 4.2, 1.7 Hz, 1H), 8.63 (s, 1H), 8.24 (dd, J = 8.4, 1.7 Hz, 1H), 7.92 (d, J = 10.0 Hz, 1H), 7.48 (dd, J = 8.3, 4.2 Hz, 1H), 7.42-7.30 (m, 2H), 6.22 (s, 1H), 4.16 (dt, J = 13.0, 6.5 Hz, 4H), 2.87 (d, J = 13.4 Hz, 1H), 2.81 (d, J = 4.7 Hz, 1H), 1.93-1.74 (m, 6H), 1.65-1.36 (m, 7H), 1.34 (s, 3H), 1.32-1.21 (m, 5H), 1.18 (s, 3H), 1.15 (s, 3H), 1.03 (s, 3H), 0.92 (s, 3H), 0.87 (s, 3H), 0.85 (s, 3H).

[0163] Example 12

[0164] Synthesis of intermediate A12

[0165]

[0166] With 6-hydroxyquinoline (2.06 mmol, 0.30 g) instead of 3-hydroxyquinoline in Example 4, and other conditions unchanged, compound A12 (white solid) 0.38 g, yield 62.9% was obtained.

[0167] ESI-MS: Br isotope characteristic peaks 292.0, 294.0 [M+H] + .

[0168] Synthesis of compound I-12

[0169]

[0170] With equal molar amount of compound A12 instead of compound A1 in the preparation method of compound I-1 in Example 1, and other conditions unchanged, the target compound I-12 (white solid) 0.06 g, yield 38.7% was obtained.

[0171] ESI-MS: 705.4 [M+H] + .

[0172] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 8.73 (dd, J = 4.2, 1.7 Hz, 1H), 8.60 (s, 1H), 8.26 (dd, J = 8.4, 1.7 Hz, 1H), 7.93 (d, J = 8.8 Hz, 1H), 7.48 (dd, J = 8.3, 4.2 Hz, 1H), 7.40 (d, J = 2.8 Hz, 1H), 7.37 (d, J = 2.5 Hz, 1H), 6.20 (s, 1H), 4.14 (dt, J = 33.9, 5.5 Hz, 4H), 2.86 (d, J = 14.2 Hz, 1H), 2.80 (d, J = 4.7 Hz, 1H), 1.82 (t, J = 7.3 Hz, 2H), 1.72 - 1.66 (m, 3H), 1.56 - 1.36 (m, 10H), 1.29 (s, 4H), 1.25 (d, J = 10.6 Hz, 5H), 1.16 (s, 3H), 1.10 (s, 3H), 0.96 (s, 3H), 0.91 (s, 3H), 0.89 (s, 3H), 0.86 (s, 3H).

[0173] Example 13

[0174] Synthesis of intermediate A13

[0175]

[0176] In a single-neck flask, 3-carboxyquinoline (1.73 mmol, 0.30 g), potassium carbonate 0.96 g (6.92 mmol), 1,2-dibromoethane 0.60 mL (6.92 mmol) were added and dissolved in 10 mL DMF, and the reaction was carried out at room temperature, and the reaction was monitored by TLC. After the reaction was completed, the organic phase was rotary evaporated and purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 6: 1 v:v) to obtain 2-bromoethyl quinoline-3-carboxylate (compound A13, white solid) 0.23 g, yield 47.6%.

[0177] ESI-MS: Bromine isotope characteristic peak 277.9, 279.9 [M+H] + .

[0178] Synthesis of compound II-1

[0179]

[0180] To a solution of 0.10 g (0.22 mmol) of CDOO in 2 mL of DMF, 0.091 g of potassium carbonate (0.66 mmol) and 0.018 g of potassium iodide (0.11 mmol) were added, stirred, 2-bromoethyl quinoline-3-carboxylate 0.184 g (0.66 mmol) was added, and the reaction was allowed to proceed at room temperature until TLC detection showed that the reaction was complete, diluted with 30 mL of ethyl acetate, washed with water 3 times, the organic layer was saturated with sodium bicarbonate 3 times, saturated with brine 3 times, and dried over anhydrous sodium sulfate overnight. The organic phase was rotary evaporated, and compound II-1 (white solid) 0.06 g was obtained by silica gel column chromatography (eluent petroleum ether: ethyl acetate = 3: 1 v:v) with a yield of 39.5%.

[0181] ESI-MS: 691.3 [M+H] + .

[0182] 1 H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.31 (d, J = 2.2 Hz, 1H), 9.01 (d, J = 2.1 Hz, 1H), 8.51 (s, 1H), 8.26 (dd, J = 8.2, 1.4 Hz, 1H), 8.16 (d, J = 8.5 Hz, 1H), 7.98 (ddd, J = 8.5, 6.9, 1.5 Hz, 1H), 7.78 (ddd, J = 8.1, 6.9, 1.2 Hz, 1H), 6.10 (s, 1H), 4.79 (ddd, J = 12.5, 7.8, 2.4 Hz, 1H), 4.58 (ddt, J = 13.0, 7.6, 4.1 Hz, 2H), 4.37 (ddd, J = 12.5, 5.4, 2.4 Hz, 1H), 2.84 (d, J = 13.3 Hz, 1H), 2.73 (d, J = 4.8 Hz, 1H), 1.93 - 1.81 (m, 1H), 1.77 - 1.55 (m, 3H), 1.52 - 1.22 (m, 11H), 1.18 (s, 3H), 1.07 (s, 3H), 0.96 (s, 6H), 0.84 (d, J = 2.2 Hz, 6H), 0.82 (s, 3H).

[0183] Example 14

[0184] Synthesis of intermediate A14

[0185]

[0186] 1,3-dibromopropane 0.72 mL (6.92 mmol) was used instead of 1,2-dibromoethane in Example 13 to obtain 3-bromopropyl quinoline-3-carboxylate (compound A14, white solid) 0.25 g with a yield of 49.3%.

[0187] ESI-MS: Bromine isotope characteristic peaks 292.0, 294.0 [M+H] + .

[0188] Synthesis of compound II-2

[0189]

[0190] Refer to the preparation method of compound II-1 in example 13, replace compound A13 with compound A14 in equimolar amount, and other conditions remain unchanged to obtain the target compound II-2 (white solid) 0.07 g, yield 45.2%.

[0191] ESI-MS: 705.3 [M+H] + .

[0192] 1 H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.35 (d, J = 2.1 Hz, 1H), 9.07 (d, J = 2.2 Hz, 1H), 8.60 (s, 1H), 8.26 (dd, J = 8.4, 1.5 Hz, 1H), 8.14 (d, J = 8.4 Hz, 1H), 7.95 (ddd, J = 8.4, 6.8, 1.5 Hz, 1H), 7.75 (ddd, J = 8.2, 6.8, 1.2 Hz, 1H), 6.17 (s, 1H), 4.45 (ddt, J = 28.5, 11.1, 5.6 Hz, 2H), 4.35 - 4.23 (m, 2H), 2.82 (d, J = 11.5 Hz, 2H), 2.17 (h, J = 6.2 Hz, 2H), 2.04 - 1.83 (m, 2H), 1.77 - 1.24 (m, 14H), 1.22 (s, 3H), 1.12 (s, 3H), 0.99 (s, 3H), 0.96 (s, 3H), 0.88 (s, 6H), 0.85 (s, 3H). Example 15

[0193] Synthesis of intermediate A15

[0194]

[0195] Refer to the preparation method of compound II-1 in example 13, replace compound A13 with compound A14 in equimolar amount, and other conditions remain unchanged to obtain the target compound II-2 (white solid) 0.07 g, yield 45.2%.

[0196] ESI-MS: Bromine isotope characteristic peaks 292.0, 294.0 [M+H] + .

[0197] Synthesis of compound II-3

[0198]

[0199] Referring to the preparation method of compound II-1 in Example 13, compound A15 was replaced with equimolar amount of compound A13, and other conditions were unchanged to obtain the target compound II-3 white solid 0.07 g, yield 44.3%.

[0200] ESI-MS: 719.4 [M+H] + .

[0201] 1 H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.32 (d, J = 2.2 Hz, 1H), 9.03 (d, J = 2.2 Hz, 1H), 8.57 (s, 1H), 8.24 (dd, J = 8.4, 1.5 Hz, 1H), 8.12 (d, J = 8.5 Hz, 1H), 7.94 (ddd, J = 8.5, 6.9, 1.5 Hz, 1H), 7.74 (ddd, J = 8.2, 6.8, 1.2 Hz, 1H), 6.17 (s, 1H), 4.40 (tt, J = 7.5, 3.7 Hz, 2H), 4.22 (dt, J = 11.6, 6.0 Hz, 1H), 4.14 (dt, J = 11.2, 6.0 Hz, 1H), 2.85 (d, J = 13.4 Hz, 1H), 2.78 (d, J = 4.8 Hz, 1H), 1.81 (dtd, J = 19.8, 9.6, 4.8 Hz, 5H), 1.54 - 1.44 (m, 4H), 1.39 (s, 1H), 1.23 (s, 7H), 1.11 (s, 8H), 0.96 (s, 3H), 0.89 (s, 3H), 0.86 (s, 3H), 0.84 (s, 6H).

[0202] Example 16

[0203] Synthesis of intermediate A16

[0204]

[0205] Referring to the preparation method of compound II-1 in Example 13, compound A15 was replaced with equimolar amount of compound A13, and other conditions were unchanged to obtain the target compound II-3 white solid 0.07 g, yield 44.3%.

[0206] ESI-MS: 719.4 [M+H] + .

[0207] Synthesis of compound II-4

[0208]

[0209] The compound II-4 (white solid) 0.06 g, 37.2% yield was obtained by referring to the preparation method of the compound II-1 in Example 13, replacing the compound A13 with the compound A16 in an equimolar amount, and other conditions unchanged.

[0210] ESI-MS: 733.4 [M+H] + .

[0211] 1 H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.31 (d, J = 2.2 Hz, 1H), 9.01 (d, J = 2.2 Hz, 1H), 8.58 (s, 1H), 8.23 (dd, J = 8.2, 1.5 Hz, 1H), 8.11-8.07 (m, 1H), 7.91 (ddd, J = 8.4, 6.9, 1.5 Hz, 1H), 7.72 (ddd, J = 8.1, 6.8, 1.2 Hz, 1H), 6.17 (s, 1H), 4.39-4.33 (m, 2H), 4.09 (q, J = 5.7 Hz, 2H), 2.84-2.74 (m, 2H), 1.77 (dd, J = 17.8, 10.4 Hz, 4H), 1.71-1.43 (m, 11H), 1.31 (s, 4H), 1.28-1.17 (m, 6H), 1.15 (s, 3H), 1.11 (s, 3H), 0.97 (s, 5H), 0.84 (s, 3H), 0.79 (s, 3H), 0.77 (s, 3H).

[0212] Example 17

[0213] Synthesis of intermediate A17

[0214]

[0215] The compound A17 (white solid) 0.37 g, 76.6% yield was obtained by replacing the 3-carboxyquinoline in Example 13 with 4-carboxyquinoline (1.73 mmol, 0.30 g), and other conditions unchanged.

[0216] ESI-MS: Bromine isotope characteristic peaks 277.9, 279.9 [M+H] + .

[0217] Synthesis of compound II-5

[0218]

[0219] The compound II-5 (white solid) 0.05 g, yield 32.9% was obtained with reference to the preparation method of the compound II-1 in example 13, replacing the compound A13 with the compound A17 in equal molar amount, and other conditions unchanged.

[0220] ESI-MS: 691.3 [M+H] + .

[0221] 1 H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.08 (d, J = 4.3 Hz, 1 H), 8.68-8.51 (m, 2 H), 8.13 (d, J = 8.4 Hz, 1 H), 7.85 (q, J = 5.6 Hz, 2 H), 7.73 (t, J = 7.8 Hz, 1 H), 6.14 (s, 1 H), 4.65 (tp, J = 9.4, 4.3 Hz, 2 H), 4.47 (dddd, J = 44.1, 12.7, 6.0, 2.9 Hz, 2 H), 2.82 (dd, J = 11.5, 6.8 Hz, 1 H), 2.72 (d, J = 4.8 Hz, 1 H), 1.91 - 1.53 (m, 4 H), 1.40 (dt, J = 29.5, 10.0 Hz, 6 H), 1.19 (s, 5 H), 1.10 (s, 5 H), 0.99 (s, 3 H), 0.91 (s, 4 H), 0.82 (s, 3 H), 0.80 (s, 6 H).

[0222] Example 18

[0223] Synthesis of intermediate A18

[0224]

[0225] The compound A18 (white solid) 0.31 g, yield 61.2% was obtained with replacing 3-carboxyquinoline with 4-carboxyquinoline (1.73 mmol, 0.30 g) in example 14, and other conditions unchanged.

[0226] ESI-MS: bromine isotope characteristic peak 292.01, 294.0 M+H] + .

[0227] Synthesis of compound II-6

[0228]

[0229] The compound II-6 (white solid) 0.05 g, yield 32.3% was obtained with reference to the preparation method of the compound II-1 in example 13, replacing the compound A13 with the compound A18 in equal molar amount, and other conditions unchanged.

[0230] ESI-MS: 705.3 [M+H] + .

[0231] 1 H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.09 (d, J = 4.4 Hz, 1H), 8.71-8.56 (m, 2H), 8.15 (d, J = 8.4 Hz, 1H), 7.99 (d, J = 4.4 Hz, 1H), 7.88 (t, J = 7.7 Hz, 1H), 7.76 (t, J = 7.7 Hz, 1H), 6.19 (s, 1H), 4.49 (h, J = 5.4 Hz, 2H), 4.26 (t, J = 6.1 Hz, 2H), 2.90-2.72 (m, 2H), 2.16 (p, J = 6.2 Hz, 2H), 1.93-1.46 (m, 10H), 1.28 (s, 5H), 1.14 (s, 6H), 1.02 (d, J = 4.0 Hz, 6H), 0.88 (s, 3H), 0.86 (s, 3H), 0.84 (s, 3H).

[0232] Example 19

[0233] Synthesis of intermediate A19

[0234]

[0235] With 4-carboxyquinoline (1.73 mmol, 0.30 g) instead of 3-carboxyquinoline in Example 15, other conditions remain the same, to obtain compound A19 (white solid) 0.27 g, yield 50.8%.

[0236] ESI-MS: Br isotope characteristic peaks 306.0, 308.0 [M+H] + .

[0237] Synthesis of compound II-7

[0238]

[0239] Referring to the preparation method of compound II-1 in Example 13, with equimolar amount of compound A19 instead of compound A13, other conditions remain the same, to obtain the target compound II-7 (white solid) 0.05 g, yield 32.3%.

[0240] ESI-MS: 719.4 [M+H] + .

[0241] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.09 (d, J = 4.5 Hz, 1 H), 8.66-8.55 (m, 2 H), 8.14 (d, J = 8.4 Hz, 1 H), 7.94 (d, J = 4.4 Hz, 1 H), 7.87 (ddd, J = 8.5, 6.8, 1.4 Hz, 1 H), 7.75 (td, J = 7.5, 1.4 Hz, 1 H), 6.18 (s, 1 H), 4.45 (t, J = 6.1 Hz, 2 H), 4.26-4.08 (m, 2 H), 2.85 (dd, J = 13.7, 4.1 Hz, 1 H), 2.77 (d, J = 4.8 Hz, 1 H), 1.93-1.44 (m, 14 H), 1.24 (s, 6 H), 1.13 (s, 3 H), 1.07 (s, 5 H), 1.01 (s, 3 H), 0.89 (s, 3 H), 0.86 (s, 3 H), 0.84 (s, 3 H).

[0242] Example 20

[0243] Synthesis of intermediate A20

[0244]

[0245] With 4-carboxyquinoline (1.73 mmol, 0.30 g) instead of 3-carboxyquinoline in Example 16, and other conditions unchanged, compound A20 (white solid) 0.27 g, yield 48.6% was obtained.

[0246] ESI-MS: characteristic peaks of bromine isotope 320.0, 322.0 [M+H] + .

[0247] Synthesis of compound II-8

[0248]

[0249] With compound A20 instead of compound A13 in equal molar amount according to the preparation method of compound II-1 in Example 13, and other conditions unchanged, the target compound II-8 (white solid) 0.06 g, yield 37.2% was obtained.

[0250] ESI-MS: 733.4 [M+H] + .

[0251] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.08 (d, J = 4.4 Hz, 1H), 8.71-8.52 (m, 2H), 8.14 (d, J = 8.4 Hz, 1H), 7.95 (d, J = 4.4 Hz, 1H), 7.86 (t, J = 7.6 Hz, 1H), 7.75 (t, J = 7.7 Hz, 1H), 6.19 (s, 1H), 4.42 (t, J = 6.3 Hz, 2H), 4.10 (q, J = 6.8 Hz, 2H), 2.89-2.71 (m, 2H), 1.88-1.58 (m, 9H), 1.57-1.30 (m, 12H), 1.15 (s, 3H), 1.13 (s, 3H), 1.01 (s, 6H), 0.84 (s, 3H), 0.81 (s, 6H).

[0252] Example 21

[0253] Synthesis of intermediate A21

[0254]

[0255] With 6-carboxyquinoline (1.73 mmol, 0.30 g) instead of 3-carboxyquinoline in Example 13, and other conditions unchanged, compound A21 (white solid) 0.27 g, yield 56.0% was obtained.

[0256] ESI-MS: bromine isotope characteristic peaks 277.9, 279.9 [M+H] + .

[0257] Synthesis of compound II-9

[0258]

[0259] With compound A21 instead of compound A13 in equal molar amount according to the preparation method of compound II-1 in Example 13, and other conditions unchanged, the target compound II-9 (white solid) 0.06 g, yield 39.5% was obtained.

[0260] ESI-MS: 691.3 [M+H] + .

[0261] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.07 (dd, J = 4.2, 1.7 Hz, 1H), 8.70 (d, J = 1.9 Hz, 1H), 8.63 (dd, J = 8.3, 1.8 Hz, 1H), 8.54 (s, 1H), 8.23-8.14 (m, 2H), 7.70 (dd, J = 8.4, 4.2 Hz, 1H), 6.13 (s, 1H), 4.79-4.72 (m, 1H), 4.55 (tdd, J = 12.5, 6.4, 2.3 Hz, 2H), 4.45-4.34 (m, 1H), 2.85 (d, J = 13.5 Hz, 1H), 2.74 (d, J = 4.8 Hz, 1H), 1.93-1.82 (m, 1H), 1.77-1.57 (m, 4H), 1.40 (s, 2H), 1.30 (s, 1H), 1.25 (d, J = 10.4 Hz, 5H), 1.16 (s, 6H), 1.07 (s, 3H), 0.96 (s, 5H), 0.85 (s, 3H), 0.84 (t, J = 1.0 Hz, 3H), 0.83 (s, 3H).

[0262] Example 22

[0263] Synthesis of intermediate A22

[0264]

[0265] With 6-carboxyquinoline (1.73 mmol, 0.30 g) instead of 3-carboxyquinoline in Example 14, and other conditions unchanged, compound A22 (white solid) 0.28 g, yield 55.2% was obtained.

[0266] ESI-MS: Br isotope characteristic peak 292.0, 294.0 [M+H] + .

[0267] Synthesis of compound II-10

[0268]

[0269] With equal molar amount of compound A22 instead of compound A13 according to the preparation method of compound II-1 in Example 13, and other conditions unchanged, the target compound II-10 (white solid) 0.06 g, yield 38.7% was obtained.

[0270] ESI-MS: 705.3 [M+H] + .

[0271] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.05 (dd, J = 4.2, 1.8 Hz, 1H), 8.75 (d, J = 2.0 Hz, 1H), 8.64 (dd, J = 8.4, 1.8 Hz, 1H), 8.60 (s, 1H), 8.25 (dd, J = 8.8, 2.0 Hz, 1H), 8.15 (d, J = 8.8 Hz, 1H), 7.67 (dd, J = 8.3, 4.2 Hz, 1H), 6.17 (s, 1H), 4.50-4.34 (m, 2H), 4.28 (ddt, J = 11.1, 7.3, 4.1 Hz, 2H), 2.82 (d, J = 9.8 Hz, 2H), 2.16 (q, J = 6.2 Hz, 2H), 1.95-1.86 (m, 1H), 1.76-1.64 (m, 2H), 1.59 (d, J = 14.0 Hz, 1H), 1.40 (t, J = 6.2 Hz, 4H), 1.34 (s, 1H), 1.30 (s, 2H), 1.25 (d, J = 10.2 Hz, 4H), 1.22 (s, 3H), 1.12 (s, 3H), 0.98 (s, 3H), 0.96 (s, 3H), 0.88 (s, 6H), 0.85 (s, 3H).

[0272] Example 23

[0273] Synthesis of intermediate A23

[0274]

[0275] With 6-carboxyquinoline (1.73 mmol, 0.30 g) instead of 3-carboxyquinoline in Example 15, and other conditions unchanged, compound A23 (white solid) 0.27 g, yield 50.8% was obtained.

[0276] ESI-MS: Br isotope characteristic peaks 306.0, 308.0 [M+H] + .

[0277] Synthesis of compound II-11

[0278]

[0279] With equal molar amount of compound A23 instead of compound A13 in reference to the preparation method of compound II-1 in Example 13, and other conditions unchanged, the target compound II-11 (white solid) 0.05 g, yield 31.6% was obtained.

[0280] ESI-MS: 719.4 [M+H] + .

[0281] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.04 (dd, J = 4.2, 1.8 Hz, 1H), 8.72 (d, J = 2.0 Hz, 1H), 8.65-8.56 (m, 2H), 8.23 (dd, J = 8.8, 2.0 Hz, 1H), 8.14 (d, J = 8.8 Hz, 1H), 7.66 (dd, J = 8.3, 4.2 Hz, 1H), 6.18 (s, 1H), 4.43-4.36 (m, 2H), 4.27-4.10 (m, 2H), 2.90-2.77 (m, 2H), 1.79 (ddd, J = 23.4, 12.2, 7.3 Hz, 5H), 1.49-1.25 (m, 12H), 1.23 (s, 5H), 1.11 (s, 3H), 1.10 (s, 3H), 0.96 (s, 3H), 0.90 (s, 3H), 0.86 (s, 3H), 0.85 (s, 3H).

[0282] Example 24

[0283] Synthesis of intermediate A24

[0284]

[0285] With 6-carboxyquinoline (1.73 mmol, 0.30 g) instead of 3-carboxyquinoline in Example 16, and other conditions unchanged, compound A24 (white solid) 0.28 g, yield 50.4% was obtained.

[0286] ESI-MS: characteristic peaks of bromine isotopes 320.0, 322.0 [M+H] + .

[0287] Synthesis of compound II-12

[0288]

[0289] With compound A24 instead of compound A13 in equal molar amount according to the preparation method of compound II-1 in Example 13, and other conditions unchanged, the target compound II-12 (white solid) 0.05 g, yield 31.0% was obtained.

[0290] ESI-MS: 733.4 [M+H] + .

[0291] 1H NMR (400 MHz, DMSO, TMS), δ ppm: δ 9.04 (dd, J = 4.2, 1.7 Hz, 1H), 8.72 (d, J = 2.0 Hz, 1H), 8.62 (d, J = 9.1 Hz, 2H), 8.24 (dd, J = 8.9, 2.0 Hz, 1H), 8.14 (d, J = 8.8 Hz, 1H), 7.66 (dd, J = 8.3, 4.2 Hz, 1H), 6.20 (s, 1H), 4.36 (t, J = 6.3 Hz, 2H), 4.11 (q, J = 5.7 Hz, 2H), 2.87-2.75 (m, 2H), 1.89-1.73 (m, 4H), 1.73-1.60 (m, 4H), 1.59-1.46 (m, 6H), 1.40 (s, 1H), 1.33 (s, 3H), 1.30 (s, 1H), 1.25 (d, J = 10.7 Hz, 5H), 1.18 (s, 3H), 1.13 (s, 3H), 0.99 (s, 3H), 0.86 (s, 3H), 0.82 (s, 3H), 0.80 (s, 3H).

[0292] Example 25

[0293] (I) In vitro anti-tumor cell proliferation experiment of the compound

[0294] The anti-tumor activity of the CDDO quinoline derivatives (test compounds) of the application was tested by tetramethyl thiazole blue colorimetry (MTT method), and CDDO and CDDO-Me (bardoxolone methyl) were selected as positive control drugs.

[0295] Instruments: super-clean workbench (SW-CJ-1FD, AIRTECH, Suqing Antai), constant-temperature CO2 incubator (3111, Thermo, USA), high-pressure sterilization pot (YXO.SG41.280, Shanghai Huaxian), centrifuge (SIGMA), multi-biology microscope (IX71, Olympus, Japan), multifunctional enzyme marker (POLARstar, Omega, USA).

[0296] Reagents: DMEM, 1640 medium (KeyGEN), fetal bovine serum (GIBCO), trypsin (SIGMA), DMSO (SIGMA).

[0297] Cell lines: mouse melanoma cell line B16-F10, human breast cancer cell line MDA-MB-231, human gastric cancer cell BGC-823, human hepatoma cell HepG2; wherein the BGC-823 cells are cultured with RPMI-1640 medium, and the B16-F10, MDA-MB-231 and HepG2 cells are cultured with DMEM (high sugar) medium (all purchased from Jiangsu Keygen Biotech Co., Ltd.).

[0298] Method: the cryopreserved cell lines are recovered and cultured in a constant temperature 37℃, 5% CO2 incubator, with liquid change once a day. When they are in the exponential growth phase and in good condition, they can be plated. Add 1 mL of 0.25% trypsin digestion solution, digest for 1-2 min, and observe the cell state under a microscope. When the adherent cells are rounded and contracted, the digestion solution can be removed, 1-2 mL of medium containing 10% fetal bovine serum is added to prepare a cell suspension, and the cell suspension is inoculated on a 96-well plate at 100 μL / well, and the periphery is sealed with PBS solution. Place in a constant temperature 37℃, 5% CO2 incubator and culture for 24 h. 3 50 50 Weigh an appropriate amount of test compound, CDDO and CDDO-Me, dissolve in DMSO to prepare a stock solution with a concentration of 10 mmol / L, then dilute with medium to prepare a series of solutions with concentrations of 0.25, 0.5, 1, 2, 4 μmol / L, and set up a blank control (DMSO is diluted with medium to a solution with a concentration of 4 μmol / L), and 3 replicate wells are set for each drug concentration. Place the 96-well plate in a constant temperature 37℃, 5% CO2 incubator for normoxic culture for 48 h. TM TM Weigh an appropriate amount of test compound, CDDO and CDDO-Me, dissolve in DMSO to prepare a stock solution with a concentration of 10 mmol / L, then dilute with medium to prepare a series of solutions with concentrations of 0.05, 0.1, 0.2, 0.4, 0.8 μmol / L, and set up a blank control (DMSO is diluted with medium to a solution with a concentration of 0.8 μmol / L), and 3 replicate wells are set for each drug concentration. Place the 96-well plate in a constant temperature 37℃, 1% O2 incubator for hypoxic culture for 72 h. TM TM Add MTT reagent (5 mg / mL) to the 96-well plate, 10 μL / well, continue to incubate for 4 h. Remove the culture medium in the plate, add 150 μL DMSO to each well, and use a multifunctional enzyme label instrument to detect the absorbance value of each well at a wavelength of 570 nm, and calculate the cell inhibition rate according to the following formula. TM 5 Cell inhibition rate % = [(blank control OD value - drug administration group OD value) / blank control group OD value] x 100%. ​

[0303] 3 screening results average value is the final inhibition rate, calculate the IC of test drug 50 value (graphpad software), 3 repeated experiments results are the final IC of measured compound 50 value.

[0304] Table 1. Inhibition of tumor cells by test compounds under 21% O2

[0305]

[0306] Table 2. Inhibition of tumor cells by test compounds under 1% O2

[0307]

[0308] According to Table 1 and Table 2, compared with CDDO, the CDDO quinoline derivatives of the application have significantly enhanced inhibition on cells MDA-MB-231, BGC-823, HepG2 and B16-F10 under normoxia and hypoxia; compared with CDDO-Me, the inhibition on cells MDA-MB-231, BGC-823, HepG2 and B16-F10 under normoxia is not much different, and the inhibition under hypoxia is not much different or significantly enhanced. Among them, the CDDO quinoline derivatives of the application, especially compound I-7, have 4.75 times higher activity under hypoxia than under normoxia in melanoma cells, 13.13 times higher activity than CDDO, and 4.75 times higher activity than CDDO-Me.

[0309] (II) Immunofluorescence experiment of compound

[0310] Instrument: high content cell imaging instrument (ImageXpress Micro).

[0311] Reagent: HIF-1α primary antibody (Proteintech), Alexa Fluor 488 labeled goat anti-mouse IgG (H+L) (Biosharp), DAPI staining solution (Biosharp), Triton X-100 (triton), bovine serum albumin V (Biosharp), 4% paraformaldehyde (Biosharp).

[0312] Method: According to the method of cell digestion counting in the experiment of in vitro anti-tumor cell proliferation of compounds, collect cell suspension (1000 / mL), inoculate B16-F10 cells uniformly in 96-well plates, and culture under normal oxygen for 24 h. Set up the drug administration group and the blank control group. In the drug administration group, compound I-7 is dissolved in DMSO to prepare a stock solution with a concentration of 10 mmol / L, and then diluted with culture medium to 0.1 μmol / L. In the blank control group, DMSO is diluted with culture medium to a solution with a concentration of 4 μmol / L. Set 3 replicate wells, and place them in a constant temperature 37℃, 1% O2 incubator for low oxygen culture for 24 h. Add 100 μL of pre-cooled 4% paraformaldehyde to each well, and fix for 10 min. Then, aspirate and wash in PBS for 3 times, 5 min each time. Add 50 μL of Triton X-100 to each well, and permeabilize for about 2 min. Then, add 100 μL of PBS to wash and aspirate, and wash for 3 times, 5 min each time. Block with an appropriate blocking solution for 0.5 h (blocking solution: 3% BSA). Incubate the primary antibody, and wash in PBS for 3 times, 5 min each time. Incubate the secondary antibody: add 60 μL of secondary antibody (dilute the secondary antibody with 5% goat serum according to the instructions, generally at a ratio of 1:500) to each well, and incubate at room temperature for 0.5 h. Take out the secondary antibody (note that all steps after the secondary antibody step need to be carried out in the dark). Wash with PBS twice, 5 min each time. Perform nuclear staining: add nuclear staining reagent (DAPI, generally 1-2 min before use, depending on the specific situation). After staining, take out the nuclear reagent, wash with PBS once, and then add 1 mL of PBS to keep it moist.

[0313] As shown in Figure 1 , Figure 2 , compared with the blank control group (marked as DMSO), the fluorescence intensity of HIF-1α protein in the drug administration group (marked as I-7) is weakened, indicating that compound I-7 can reduce the content of HIF-1α protein in cells under hypoxia.

[0314] (Three) Effect of the compound on HIF-1α protein and downstream protein VEGF protein

[0315] Instrument: Tanon gel imaging system (Tanon-5200, Shanghai Tianni), Mini Trans-Blot 170-3930 (Bio-Rad).

[0316] Reagent: BCA protein concentration determination kit (KeyGEN), SDS-PAGE protein loading buffer (5X) (KeyGEN), BeyoColor TM color pre-stained protein molecular weight marker (10-170kD) (Bi Yun Tian, product number: P0075), BeyoGel TMSDS-PAGE precast gel (Tris-Gly, 4-20%, 12 well) (Bi Yun Tian, item number: P0057A), SDS-PAGE running buffer (Tris-Gly, powder) (Bi Yun Tian, item number: P0014B), PVDF membrane (imported, 6.6 x 8.5 cm, 0.45 μm) (Bi Yun Tian, item number: FFP32), Western transfer buffer (Bi Yun Tian, item number: P0021B), QuickBlock TM Western blocking buffer (Bi Yun Tian, item number: P0252), QuickBlock TM Western primary antibody diluent (Bi Yun Tian, item number: P0256), QuickBlock TM Western secondary antibody diluent (Bi Yun Tian, item number: P0258), Western wash buffer (10X) (Bi Yun Tian, item number: P0023C3), recombinant Anti-HIF-1α antibody (Abeam), recombinant Anti-Vegf antibody (Abeam).

[0317] Method: B16-F10 cells in logarithmic growth phase were inoculated in 10 cm culture dishes (2 x 10 5The cells are cultured for 24 hours at 37°C in a 5% CO2 incubator, and then compound I-7 is added to the designated dishes (dissolved in DMSO to prepare a mother liquor with a concentration of 10 mmol / L, and then diluted with a culture medium to 0.05, 0.1 and 0.2 μmol / L), and two blank control groups (DMSO is diluted with a culture medium to a solution with a concentration of 4 μmol / L) are set. One of the blank control groups is continuously cultured for 48 hours under normal oxygen, and the other blank control group and the drug administration group are continuously cultured for 48 hours at 37°C under 1% O2, and then the cells are collected, centrifuged, washed with PBS, and 100 μL of IP lysis buffer (containing protease inhibitors) is added for lysis at 0°C for 1 hour. The supernatant is discarded after centrifugation at 4°C and 10000g for 5 minutes, and the protein sample is quantified using a BCA protein concentration determination kit, and then stored at -80°C for use. The protein sample is diluted to 3 μg / μL using 5X SDS-PAGE protein loading buffer and lysis buffer. After cooling to room temperature, the sample and Marker are added to an SDS-PAGE gel, and the power is turned on at 60V. After the sample is electrophoresed into the separation gel, the voltage is adjusted to 90V, and the target band is eluted to about 2 / 3 of the gel according to the Marker, and the electrophoresis is terminated. The separation gel containing the target band is cut and placed on a pre-activated PVDF membrane, and a mark is made, and the membrane is transferred at a constant current of 300mA for 60 minutes. The transferred membrane is placed in a dish containing blocking solution, and slowly shaken and incubated for 1 hour, and then the blocking solution is discarded. The primary antibody diluted with a primary antibody diluent is added, and incubated at 4°C overnight. After recovering the primary antibody, the membrane is washed with Western washing solution for 3 times, each for 10 minutes. The secondary antibody diluted with a secondary antibody diluent is added, and slowly shaken and incubated at 4°C for 1.5 hours, and then the membrane is washed with Western washing solution for 3 times, each for 8 minutes. The PVDF membrane is taken out, and an appropriate amount of prepared working solution is added, and covered with plastic wrap. The gel is developed and imaged by a Tianen gel imaging system, and gray scale analysis is performed by Image J software.

[0318] As shown in Figure 3 , Figure 4 , Figure 5 Under hypoxia, the content of HIF-1α protein is increased, the content of HIF-1α is reduced with the increase of the concentration of the compound, and the downstream protein VEGF of HIF-1α is also reduced in a concentration-dependent manner.

[0319] In summary, the CDDO quinoline derivative compound of the present application can inhibit the proliferation of the above-mentioned tumors under hypoxia, and reduce the protein content of HIF-1α and VEGF, and is expected to become a new anti-tumor candidate drug, and is worthy of further research.

Claims

1. CDDO quinoline derivatives with structures as shown in Formula III: ； in, R is selected from , R1 is selected from , , R2 is selected from , , n is an integer selected from 2 to 5.

2. The CDDO quinoline derivative according to claim 1, characterized in that: R is selected from , R1 is selected from , , R2 is selected from , , ; n is selected from integers between 2 and 5; but does not include: R is selected from R1 is selected from n=5.

3. The CDDO quinoline derivative according to claim 2, characterized in that: R is selected from , R1 is selected from , , R2 is selected from , , ; n is selected from integers between 2 and 5; but does not include: R is selected from R1 is selected from , n=5.

4. A CDDO quinoline derivative, characterized in that: Selected from compounds with the following structures: 。 5. CDDO quinoline derivatives with the following structures: 。 6. A method for preparing the CDDO derivative according to claim 1, characterized in that: When R is selected The synthesis route is as follows: ； R1 and n are as described in claim 1 above; in: Step (1): Using potassium carbonate as a catalyst and N,N-dimethylformamide as a reaction solvent, the quinoline hydroxyl compound represented by formula R1-OH reacts with... The dibromoalkane shown reacts at room temperature to give intermediate d; Step (2): Using potassium carbonate and potassium iodide as catalysts and N,N-dimethylformamide as reaction solvent, intermediate d reacts with CDDO at room temperature to obtain CDDO quinoline derivative; When R is selected The synthesis route is as follows: ； R1, n are as described in claim 1; in: Step (1): Using potassium carbonate as a catalyst and N,N-dimethylformamide as a reaction solvent, the formula is... The quinoline hydroxy compounds shown are similar to those in formula [formula missing]. The dibromoalkane shown was reacted at room temperature to give intermediate h; wherein the molar ratio of the quinoline carboxyl compound to the dibromoalkane was 1:3 to 1:5; and the volume ratio of the dibromoalkane to potassium carbonate was 1:1 to 1:1.

5. Step (2): Using potassium carbonate and potassium iodide as catalysts and N,N-dimethylformamide as reaction solvent, intermediate d reacts with CDDO at room temperature to obtain CDDO quinoline derivative; wherein, the molar ratio of CDDO to intermediate h is 1:1.1 to 1:1.3; the molar ratio of CDDO to potassium carbonate is 1:3 to 1:5; and the molar ratio of CDDO to potassium iodide is 1:0.3 to 1:0.

5.

7. The method for preparing the CDDO derivative according to claim 6, characterized in that: When R is selected hour, In step (1), the molar ratio of the quinoline hydroxyl compound to the dibromoalkane is 1:3 to 1:5; the volume ratio of the dibromoalkane to potassium carbonate is 1:1 to 1:1.

5. In step (2), the molar ratio of CDDO to intermediate d is 1:1.1 to 1:1.5; the molar ratio of CDDO to potassium carbonate is 1:4 to 1:6; and the molar ratio of CDDO to potassium iodide is 1:0.3 to 1:0.

5. When R is selected hour, In step (1), the molar ratio of the quinoline carboxyl compound to the dibromoalkane is 1:3 to 1:5; the volume ratio of the dibromoalkane to potassium carbonate is 1:1 to 1:1.

5. In step (2), the molar ratio of CDDO to intermediate h is 1:1.1 to 1:1.3; the molar ratio of CDDO to potassium carbonate is 1:3 to 1:5; and the molar ratio of CDDO to potassium iodide is 1:0.3 to 1:0.

5.

8. The use of the CDDO quinoline derivative according to any one of claims 1-5 in the preparation of antitumor drugs.

9. The use of the CDDO quinoline derivative according to any one of claims 1-5 in the preparation of an antitumor drug by inhibiting HIF-1α protein.

10. The application according to claim 8, characterized in that: The tumors mentioned are melanoma, liver cancer, breast cancer, and stomach cancer.

11. A pharmaceutical composition, characterized in that: It uses the CDDO quinoline derivative as described in any one of claims 1-5 as the active ingredient, supplemented with a pharmaceutically acceptable carrier, to formulate any pharmaceutically acceptable dosage form.

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