Alpha, beta unsaturated ketone derivative as well as preparation method and application thereof

Abstract

The invention belongs to the field of medicinal chemistry, and discloses an alpha, beta unsaturated ketone derivative as well as a preparation method and application thereof. The structural general formula of the alpha, beta unsaturated ketone derivative disclosed by the invention is shown as a formula (I): # imgabs0 #, in the formula (I), R1 is H,-OR4,-COOR4,-OCOR4,-NHCOR4 or-CONHR4, and R1 can be monosubstituted or disubstituted; r4 is alkyl of H and C1-C3; r2 is a substituted aromatic ring or a substituted aromatic heterocyclic ring; and R3 is H, Me, Et or CN. The compound interferes with microtubule dynamic balance, has proliferation inhibition activity on tumor cells, blocks the tumor cells in a G2 / M phase and induces cell apoptosis, and can be used for preparing medicines for treating malignant tumors caused by microtubule dynamic balance imbalance, especially tumors resistant to chemotherapeutics.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and particularly to an α,β-unsaturated ketone derivative. Background Art

[0002] The latest global cancer burden data released by the International Agency for Research on Cancer (IARC) shows that the number of new cancer cases worldwide reached 20 million in 2022, and the number of death cases was approximately 9.7 million (CA Cancer J. Clin. 2024, 74(3): 229 - 263). Chemotherapy is one of the main means of cancer treatment, but the clinical application of chemotherapeutic drugs is limited due to drug resistance and side effects. Therefore, it is urgent to develop safe and effective anti-tumor drugs.

[0003] Microtubules are hollow cylindrical structures formed by the polymerization of α-tubulin and β-tubulin. Under normal physiological conditions, microtubules are in a dynamic equilibrium state. As an important component of the cytoskeleton, microtubules are related to various biological functions such as the transport of intracellular substances, cell movement, maintenance of cell morphology, and cell proliferation (Biochim. Biophys. Acta, Rev. Cancer. 2022, 1877(5): 188777). During cell proliferation, microtubules are involved in the formation of the spindle and are closely related to the distribution of genetic material. Tumor cells have the ability of rapid proliferation, and their mitosis process is frequent and the cell cycle is shorter than that of normal cells. Therefore, microtubules in tumor cells are in an abnormally activated state. Microtubule-targeting agents (MTAs) bind to microtubules, promote or inhibit microtubule polymerization, disrupt the dynamic equilibrium of microtubules, interfere with cell mitosis, and induce the death of tumor cells (Eur. J. Med. Chem. 2024, 274: 116543). Previously, the present inventors reported the anti-tumor effect of ureido compounds (CN114276303A). This type of compound can induce ferroptosis, but has tumor selectivity and can only induce ferroptosis in human colorectal cancer, and has poor inhibitory effects on other types of tumors such as breast cancer and fibrosarcoma. Summary of the Invention

[0004] In order to solve the above technical problems, the present invention discloses an α,β-unsaturated ketone derivative, and provides a specific preparation method of this type of compound and its pharmaceutical application as a microtubule polymerization inhibitor.

[0005] To achieve the above object, the technical solution of the present invention is realized as follows:

[0006] An α,β-unsaturated ketone derivative, the general structural formula of which is shown in formula (I):

[0007]

[0008] Among them, R 1 is H, -OR 4 , -COOR 4 , -OCOR 4 , -NHCOR 4 or -CONHR 4 , R 1 can be mono-substituted or di-substituted; R 4 is H, an alkyl group of C1-C3; R 2 is H, a substituted aromatic ring or a substituted heteroaromatic ring; R 3 is H, Me, Et or CN.

[0009] Furthermore, the above R 1 is H, -OMe, -OEt, -COOMe, -COOEt, -CONH 2 , -CONHMe, -NHCOMe or -NHCOEt. R 2 is Among them, R 5 is H, F, CH 3 , t-Bu, CF 3 , CN, OH, OCH 3 , NO 2 , NH 2 , NHCH 3 , N(CH 3 ) 2 or CONH 2 , R 5 can be mono-substituted, di-substituted or tri-substituted.

[0010] Furthermore, the above R 1 is H, -OMe, -OEt, -COOMe, -COOEt, -CONH 2 or -CONHMe.

[0011] The preparation method of the above α, β-unsaturated ketone derivative is as follows:

[0012]

[0013] Among them, R 1 is H, -OR 4 , -COOR 4 , -OCOR 4 , -NHCOR 4 or -CONHR 4 , R 1 can be mono-substituted or di-substituted; R 4 is H, an alkyl group of C1-C3; R 2is a substituted aromatic ring or a substituted heteroaromatic ring; R 3 is H, Me, Et or CN.

[0014] The specific preparation steps are as follows:

[0015] (1) Compound II is dissolved in anhydrous dichloromethane and reacts with di-tert-butyl dicarbonate under the action of a base to obtain Compound III; wherein, the structural formula of Compound II is The structural formula of Compound III is

[0016] (2) Compound III is dissolved in methanol, and the benzyl group is removed under the action of hydrogen using Pd / C or Raney Ni as a catalyst to obtain Compound IV; the structural formula of Compound IV is

[0017] (3) Compound IV is dissolved in anhydrous dichloromethane and reacts with an acyl chloride under the action of a base to form Compound V; wherein, the structural formula of Compound V is

[0018] (4) Compound V is dissolved in anhydrous dichloromethane, and the Boc protecting group is removed with trifluoroacetic acid to obtain Compound I.

[0019] The above base is triethylamine or DIPEA; the acyl chloride is

[0020] wherein, R 3 is H, Me, Et or CN; R 5 is H, F, CH 3 , t-Bu, CF 3 , CN, OH, OCH 3 , NO 2 , NH 2 , NHCH 3 , N(CH 3 ) 2 or CONH 2 , and R 5 can be mono-substituted, di-substituted or tri-substituted.

[0021] A pharmaceutically acceptable salt of an α,β-unsaturated ketone derivative, which is an acid addition salt of a compound of general formula (I), wherein the acid used for salification is hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or ferulic acid.

[0022] The present invention also discloses a pharmaceutical composition comprising an α,β-unsaturated ketone derivative or a pharmaceutically acceptable salt thereof. This pharmaceutical composition can be formulated into common pharmaceutical preparations, such as tablets, capsules, syrups, suspensions, injections, by adding pharmaceutically acceptable carriers, and common pharmaceutical excipients, such as flavors, sweeteners, liquid or solid fillers or diluents, can be added.

[0023] Use of the α,β-unsaturated ketone derivative or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition of the present invention in the preparation of a microtubule polymerization inhibitor drug. Use of the α,β-unsaturated ketone derivative of the present invention and its stereoisomers, hydrates, solvates or crystals in the preparation of a microtubule polymerization inhibitor drug is also within the scope of protection of the present invention.

[0024] The above-mentioned microtubule polymerization inhibitor is a drug for treating malignant tumors.

[0025] Furthermore, the above-mentioned malignant tumors are breast cancer, cervical cancer, fibrosarcoma, ovarian cancer, liver cancer, lung cancer, colon cancer, prostate cancer or chronic myeloid leukemia.

[0026] The beneficial effects produced by the present invention are as follows:

[0027] (1) The present invention discloses an α,β-unsaturated ketone derivative represented by the general formula (I). The pharmacological experimental results show that the α,β-unsaturated ketone compound (I) of the present invention has proliferation inhibitory activity against tumor cells, blocks tumor cells in the G2 / M phase, induces apoptosis, and can be used for the preparation of a drug for treating malignant tumors caused by the imbalance of tubulin homeostasis; the present invention also discloses a preparation method of the α,β-unsaturated ketone derivative.

[0028] (2) The compounds in the present invention show better inhibitory activity against breast cancer, fibrosarcoma, cervical cancer, etc., and also show strong inhibitory activity against doxorubicin-resistant breast cancer cells, which is superior to the positive control colchicine, indicating that the compounds of the present invention can be used for the treatment of tumors resistant to chemotherapy drugs.

[0029] (3) The α,β-unsaturated ketone derivative prepared by the present invention can also form a covalent interaction with tubulin, enhance the binding ability with tubulin, disrupt the dynamic balance of microtubules, inhibit the function of drug efflux proteins, and has an inhibitory effect on drug-resistant tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art.

[0031] Figure 1Compound Ⅰ-1 arrests HT-1080 cells at the G2 / M phase. Among them, A) Flow cytometry was used to test the effect of different concentrations of Compound Ⅰ-1 on the cell cycle; B) Statistical chart of the number of G2 / M phase cells under treatment with different concentrations of Compound Ⅰ-1.

[0032] Figure 2 Compound Ⅰ-1 induces apoptosis in HT-1080 cells. Among them, A) Flow cytometry was used to test the effect of different concentrations of Compound Ⅰ-1 on apoptosis; B) Statistical chart of the number of apoptotic cells under treatment with different concentrations of Compound Ⅰ-1.

[0033] Figure 3 Effect of different concentrations of Compound Ⅰ-1 on colony formation of HT-1080 cells.

[0034] Figure 4 Effect of different concentrations of Compound Ⅰ-1 on tubulin polymerization.

[0035] Figure 5 Binding mode of the compound to tubulin; among them, N: cell viability before washing; W: cell viability after washing. Detailed implementation mode

[0036] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention, but the protection scope of the present invention is not limited thereto.

[0037] Example 1

[0038] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0039] (1) Synthesis of tert-butyl 4-benzyl-6-methoxy-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅲ-1)

[0040] Dissolve Ⅱ-1 (300 mg, 1.18 mmol), (Boc) 2 O (514 mg, 2.36 mmol), and triethylamine (238 mg, 2.36 mmol) in DCM (20 mL), and reflux and stir at 40 °C. Monitor by TLC (PE:EtOAc = 2:1). After 22 h, the reaction is complete, stop the reaction, cool to room temperature, wash with water (50 mL × 3) and saturated NaCl (50 mL × 3), and dry with anhydrous Na 2 SO 4 Dry. Column chromatography (PE:EtOAc = 50:1) gives 303 mg of white solid, with a yield of 72%. 1 H NMR (300 MHz, CDCl 3)δ 7.38 - 7.31 (m, 2H), 7.29 - 7.24 (m, 3H), 7.24 - 7.20 (m, 1H), 6.25 - 6.17 (m, 2H), 4.50 (s, 2H), 3.81 (t, J = 4.9 Hz, 2H), 3.68 (s, 3H), 3.42 (t, J = 5.0 Hz, 2H), 1.52 (s, 9H).

[0041] (2) Synthesis of tert - butyl 6 - methoxy - 3,4 - dihydroquinoxaline - 1(2H) - carboxylate (IV - 1)

[0042] Dissolve III - 1 (1.00 g, 2.82 mmol) in MeOH (15 mL), add Pd / C (60 mg, 0.56 mmol) in portions, and stir under reflux at 65 °C. Monitor by TLC (PE:EtOAc = 5:1). After 6 h, the reaction is complete. Stop the reaction, filter by suction, evaporate the solvent under reduced pressure, and triturate with PE (8 mL) to obtain 640 mg of white solid powder, with a yield of 85.79%. 1 H NMR (300 MHz, CDCl 3 )δ 7.37 (br, 1H), 6.24 (dd, J = 2.3, 8.9 Hz, 1H), 6.10 (d, J = 2.4 Hz, 1H), 3.99 (s, 1H), 3.76 - 3.71 (m, 5H), 3.39 (t, J = 4.8 Hz, 2H), 1.51 (s, 9H).

[0043] (3) Synthesis of tert - butyl 6 - methoxy - 4 - [3 - (3 - methoxyphenyl) - 1 - oxoprop - 2 - en - 1 - yl] - 1,2,3,4 - tetrahydroquinoxaline - 1 - carboxylate (V - 1)

[0044] Dissolve 3 - methoxycinnamic acid (303 mg, 1.70 mmol) and DMF (5 drops) in anhydrous DCM, stir in an ice bath for 30 min, then slowly add the DCM solution of oxalyl chloride (288 mg, 2.27 mmol), stir at room temperature for 7 h, and concentrate under reduced pressure. Dissolve the reaction mixture in anhydrous DCM. Dissolve IV - 1 (300 mg, 1.14 mmol), DIPEA (293 mg, 2.27 mmol), and DMAP (27 mg, 0.23 mmol) in anhydrous DCM, and stir in an ice bath for 30 min. Slowly add the anhydrous DCM solution of 3 - (3 - methoxyphenyl)acryloyl chloride at 0 °C, and stir at room temperature for 5 h. Monitor by TLC (PE:EtOAc = 2:1). After the reaction is complete, dilute with DCM (30 mL), wash with 1N HCl (30 mL × 3), water (30 mL × 3), 1N NaOH (30 mL × 3), and saturated NaCl (30 mL × 3), and anhydrous Na 2 SO 4Dry overnight. Filter and concentrate to obtain the crude product, and purify by column chromatography (PE:EtOAc = 8:1 - 6:1) to obtain 320 mg of yellow oil, with a yield of 66%. 1 H NMR(300MHz,CDCl 3 )δ7.80(d,J = 8.6Hz,1H),7.73(d,J = 15.6Hz,1H),7.29(d,J = 7.9Hz,1H),7.08(d,J = 7.6Hz,1H),7.01 - 6.94(m,2H),6.90(dd,J = 1.8,8.2Hz,1H),6.78(dd,J = 2.8,9.1Hz,1H),6.69(d,J = 2.4Hz,1H),4.03(t,J = 6.1Hz,2H),3.85(t,J = 6.3Hz,2H),3.81(s,3H),3.73(s,3H),1.54(s,9H).

[0045] (4) Synthesis of 3-(3-methoxyphenyl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-1)

[0046] Dissolve Ⅴ-1 (500 mg, 1.18 mmol) in DCM (6 mL) solution, add TFA (3 mL), and stir at room temperature. After the reaction is complete, concentrate under reduced pressure, dissolve in ethyl acetate (30 mL), and wash with saturated NaHCO 3 (30 mL×3), water (30 mL×3), saturated NaCl (30 mL×3), and dry over anhydrous Na 2 SO 4 Filter and concentrate to obtain the crude product. Purify by column chromatography (PE:EtOAc = 6:1) to obtain 165 mg of yellow solid, which is compound Ⅰ-1, with a yield of 43%. 1 H NMR(300MHz,CDCl 3 )δ7.71(d,J = 15.7Hz,1H),7.29 - 7.24(m,2H),7.09(d,J = 8.2Hz,1H),7.02(d,J = 7.3Hz,1H),6.93 - 6.86(m,1H),6.71 - 6.64(m,2H),6.60(d,J = 9.4Hz,1H),3.98(t,J = 5.0Hz,2H),3.81(s,3H),3.68(s,3H),3.47(t,J = 5.1Hz,2H). 13 C NMR(75MHz,CDCl 3)δ164.8,159.9,151.2,142.2,136.6,132.0,129.9,124.8,120.4,120.1,115.7,115.4,113.2,113.1,110.4,56.0,55.3,42.9,39.9.ESI-HRMS(m / z):calcd.for C 19 H 20 N 2 NaO 3 [M+Na] + :347.13661;found:347.13662.

[0047]

[0048] Example 2

[0049] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0050] The preparation methods of steps (1) and (2) are the same as those in Example 1.

[0051] (3) Synthesis of tert-butyl 6-methoxy-4-[3-(4-methoxyphenyl)-1-oxoprop-2-enyl]-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-2)

[0052] Using 4-methoxycinnamic acid (303 mg, 1.70 mmol) as the raw material, the operation process is the same as that of compound Ⅴ-1, and 197 mg of yellow solid is obtained, with a yield of 41%. 1 H NMR(300MHz,CDCl3)δ7.85-7.77(m,1H),7.73(d,J=15.5Hz,1H),7.44(d,J=8.6Hz,2H),6.92-6.83(m,3H),6.78(dd,J=2.8,9.1Hz,1H),6.71(d,J=2.5Hz,1H),4.03(t,J=6.1Hz,2H),3.88-3.81(m,5H),3.74(s,3H),1.53(s,9H).

[0053] (4) Synthesis of 3-(4-methoxyphenyl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-2)

[0054] Using Ⅴ-2 (200 mg, 0.47 mmol) as the raw material, the operation process is the same as that of the target compound Ⅰ-1, and 68 mg of yellow solid is obtained, with a yield of 45%, which is compound Ⅰ-2. 11H NMR (300 MHz, CDCl3) δ 7.71 (d, J = 15.5 Hz, 1H), 7.43 (d, J = 8.3 Hz, 2H), 6.95 (d, J = 15.5 Hz, 1H), 6.87 (d, J = 8.3 Hz, 2H), 6.72 - 6.62 (m, 2H), 6.59 (d, J = 8.2 Hz, 1H), 3.96 (t, J = 5.0 Hz, 2H), 3.82 (s, 3H), 3.68 (s, 3H), 3.45 (t, J = 4.8 Hz, 2H). 13 13C NMR (75 MHz, CDCl3) δ 165.2, 161.0, 151.2, 142.0, 132.0, 130.0, 128.0, 125.1, 117.2, 115.7, 114.3, 113.0, 110.4, 56.0, 55.4, 42.9, 39.8. ESI - HRMS (m / z): calcd. for C19H20N2NaO3 [M + Na]+: 347.13661; found: 347.13657.

[0055]

[0056] Example 3

[0057] The preparation method of the α,β - unsaturated ketone derivative in this example is as follows:

[0058] The preparation methods of steps (1) and (2) are the same as those in Example 1.

[0059] (3) Synthesis of tert - butyl 6 - methoxy - 4 - [3 - (4 - fluorophenyl) - 1 - oxoprop - 2 - en - 1 - yl] - 1,2,3,4 - tetrahydroquinoxaline - 1 - carboxylate (Ⅴ - 3)

[0060] Using 4 - fluorocinnamic acid (200 mg, 1.20 mmol) as the raw material, the operation process is the same as that of compound Ⅴ - 1, and 139 mg of yellow solid is obtained, with a yield of 28%. 1 1H NMR (300 MHz, CDCl3) δ 7.81 (d, J = 9.0 Hz, 1H), 7.73 (d, J = 15.6 Hz, 1H), 7.51 - 7.42 (m, 2H), 7.09 - 7.01 (m, 2H), 6.91 (d, J = 15.6 Hz, 1H), 6.79 (dd, J = 1.9, 9.0 Hz, 1H), 6.71 - 6.65 (m, 1H), 4.03 (t, J = 6.2 Hz, 2H), 3.85 (t, J = 6.3 Hz, 2H), 3.74 (s, 3H), 1.53 (s, 9H).

[0061] (4) Synthesis of 3-(4-Fluorophenyl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-3)

[0062] Using Ⅴ-3 (100 mg, 0.24 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ-1, and 50 mg of yellow solid was obtained with a yield of 66%, namely compound Ⅰ-3. 1 H NMR (300 MHz, CDCl 3 +D 2 O) δ 7.71 (d, J = 15.6 Hz, 1H), 7.52 - 7.41 (m, 2H), 7.10 - 6.94 (m, 3H), 6.71 - 6.56 (m, 3H), 3.97 (t, J = 4.8 Hz, 2H), 3.68 (s, 3H), 3.46 (t, J = 4.8 Hz, 2H). 13 C NMR (75 MHz, CDCl 3 ) δ 164.7, 163.6 (d, 1 J C-F = 248.8 Hz), 151.2, 141.0, 132.0, 131.5 (d, 4 J C-F = 3.2 Hz), 129.7 (d, 3 J C-F = 8.3 Hz), 124.8, 119.4 (d, 5 J C-F = 2.0 Hz), 116.0 (d, 2 J C-F = 21.7 Hz), 115.7, 113.0, 110.5, 56.0, 42.9, 39.8. ESI-HRMS (m / z): calcd. for C 18 H 17 FN 2 NaO 2 [M+Na] + : 335.11663; found: 335.11669.

[0063]

[0064] Example 4

[0065] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0066] The preparation methods of steps (1) and (2) are the same as those in Example 1.

[0067] (3) Synthesis of tert-butyl 6-methoxy-4-[3-(4-cyanophenyl)-1-oxoprop-2-en-1-yl]-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-4)

[0068] Using 4-cyanocinnamic acid (200 mg, 1.15 mmol) as the raw material, the operation process was the same as that of compound Ⅴ-1, and 230 mg of yellow solid was obtained with a yield of 47%. 1 H NMR (300 MHz, CDCl 3 ) δ 7.82 (d, J = 9.3 Hz, 1H), 7.74 (d, J = 15.6 Hz, 1H), 7.65 (d, J = 8.2 Hz, 2H), 7.55 (d, J = 8.1 Hz, 2H), 7.06 (d, J = 15.6 Hz, 1H), 6.80 (dd, J = 2.6, 9.2 Hz, 1H), 6.67 - 6.61 (m, 1H), 4.04 (t, J = 6.3 Hz, 2H), 3.85 (t, J = 6.2 Hz, 2H), 3.74 (s, 3H), 1.54 (s, 9H).

[0069] (4) Synthesis of 3-(4-cyanophenyl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-4)

[0070] Using Ⅴ-4 (230 mg, 0.55 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ-1, and 108 mg of yellow solid was obtained with a yield of 62%, which was compound Ⅰ-4. 1 H NMR (300 MHz, CDCl 3 ) δ 7.72 (d, J = 15.5 Hz, 1H), 7.64 (d, J = 8.2 Hz, 2H), 7.56 (d, J = 8.0 Hz, 2H), 7.14 (d, J = 15.6 Hz, 1H), 6.73 - 6.65 (m, 1H), 6.64 - 6.54 (m, 2H), 3.97 (t, J = 5.3 Hz, 2H), 3.68 (s, 3H), 3.48 (t, J = 5.3 Hz, 2H). 13 C NMR (75 MHz, CDCl 3 ) δ 163.8, 151.2, 139.8, 139.6, 132.7, 132.1, 128.2, 124.4, 123.3, 118.6, 115.7, 113.1, 112.7, 110.5, 56.0, 42.8, 39.8. ESI-HRMS (m / z): calcd. for C 19 H 17 N 3 NaO 2 [M+Na]+ : 342.12130; found: 342.12430.

[0071]

[0072] Example 5

[0073] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0074] The preparation methods of steps (1) and (2) are the same as those in Example 1.

[0075] (3) Synthesis of tert-butyl 6-methoxy-4-[3-(4-nitrophenyl)-1-oxoprop-2-en-1-yl]-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-5)

[0076] Using 4-nitrocinnamic acid (200 mg, 1.04 mmol) as the raw material, the operation process is the same as that of compound Ⅴ-1, and 299 mg of yellow solid is obtained, with a yield of 66%. 1 H NMR (300 MHz, CDCl 3 ) δ 8.23 (d, J = 8.6 Hz, 2H), 7.88 - 7.72 (m, 2H), 7.62 (d, J = 8.6 Hz, 2H), 7.11 (d, J = 15.6 Hz, 1H), 6.81 (dd, J = 2.5, 9.1 Hz, 1H), 6.71 - 6.60 (m, 1H), 4.05 (t, J = 5.9 Hz, 2H), 3.86 (t, J = 6.1 Hz, 2H), 3.75 (s, 3H), 1.54 (s, 9H).

[0077] (4) Synthesis of 3-(4-nitrophenyl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-5)

[0078] Using Ⅴ-5 (240 mg, 0.55 mmol) as the raw material, the operation process is the same as that of the target compound Ⅰ-1, and 160 mg of red solid is obtained, with a yield of 86%, which is compound Ⅰ-5. 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.34 - 8.14 (m, 2H), 8.02 - 7.79 (m, 2H), 7.74 - 7.59 (m, 1H), 7.31 (br, 1H), 6.70 - 6.54 (m, 2H), 5.83 (br, 1H), 3.81 (br, 2H), 3.65 - 3.50 (m, 3H), 3.35 (s, 2H). 13 C NMR (75 MHz, DMSO-d 6)δ168.3,154.8,152.8,146.7,143.8,138.1,134.1,129.8,129.3,128.5,120.3,118.2,115.1,60.7,47.0.ESI-HRMS(m / z):calcd.forC 18 H 17 N 3 NaO 4 [M+Na] + :362.11113;found:362.11115.

[0079]

[0080] Example 6

[0081] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0082] The preparation methods of steps (1)-(4) are the same as those in Example 5.

[0083] (5) Synthesis of 3-(4-aminophenyl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-6) Fe (26 mg, 0.47 mmol) and 0.15 mol / L HCl (314 μL, 0.047 mmol) were respectively added to the ethanol solution containing compound Ⅰ-5 (80 mg, 0.24 mmol), refluxed for 5 h, filtered, concentrated, dissolved in DCM, washed with water (30 mL×3), saturated NaHCO 3 (30 mL×3), saturated NaCl (30 mL×3), dried over anhydrous Na 2 SO 4 dried, filtered by suction, concentrated, and the crude product was purified by column chromatography (PE:EtOAc = 3:1) to obtain 53 mg of yellow solid with a yield of 73%, namely compound Ⅰ-6. 1 H NMR (300 MHz, CDCl 3 )δ7.70 (d, J = 15.5 Hz, 1H), 7.34 (d, J = 8.0 Hz, 2H), 6.91 (d, J = 15.4 Hz, 1H), 6.75 (s, 1H), 6.72 - 6.59 (m, 4H), 4.06 - 3.89 (m, 4H), 3.72 (s, 3H), 3.53 - 3.42 (m, 2H). 13 C NMR (75 MHz, CDCl 3) δ 165.6, 151.1, 148.3, 142.7, 131.9, 129.7, 125.5, 125.2, 115.7, 115.2, 114.9, 113.0, 110.4, 56.0, 42.9, 39.8. ESI-HRMS (m / z): calcd. for C 18 H 19 N 3 NaO 2 [M + Na] + : 332.13695; found: 332.13673.

[0084]

[0085] Example 7

[0086] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0087] The preparation methods of steps (1)-(2) are the same as those in Example 1.

[0088] (3) Synthesis of tert-butyl 6-methoxy-4-[3-(3,4-dimethoxyphenyl)-1-oxoprop-2-enyl]-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-6)

[0089] Using 3,4-dimethoxycinnamic acid (200 mg, 0.96 mmol) as the raw material, the operation process is the same as that of compound Ⅴ-1, and 137 mg of light yellow solid is obtained, with a yield of 31%. 1 H NMR (300 MHz, CDCl 3 ) δ 7.81 (d, J = 9.2 Hz, 1H), 7.71 (d, J = 15.5 Hz, 1H), 7.08 (d, J = 8.2 Hz, 1H), 6.98 (s, 1H), 6.87 (d, J = 4.2 Hz, 1H), 6.85 - 6.81 (m, 1H), 6.77 (d, J = 9.3 Hz, 1H), 6.74 - 6.69 (m, 1H), 4.03 (t, J = 5.8 Hz, 2H), 3.90 (s, 3H), 3.87 (s, 3H), 3.86 - 3.80 (m, 2H), 3.73 (s, 3H), 1.54 (s, 9H).

[0090] (4) Synthesis of 3-(3,4-dimethoxyphenyl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-7)

[0091] Using Ⅴ-6 (274 mg, 0.60 mmol) as the raw material, the operation process was the same as that of target compound Ⅰ-1, and 150 mg of yellow solid was obtained with a yield of 39%, namely compound Ⅰ-7. 1 H NMR (300 MHz, CDCl 3 ) δ 7.69 (d, J = 15.2 Hz, 1H), 7.09 (d, J = 7.7 Hz, 1H), 7.03 - 6.90 (m, 2H), 6.85 (d, J = 7.8 Hz, 1H), 6.75 - 6.63 (m, 2H), 6.60 (d, J = 8.2 Hz, 1H), 4.02 - 3.94 (m, 2H), 3.90 (s, 3H), 3.87 (s, 3H), 3.68 (s, 3H), 3.51 - 3.40 (m, 2H). 13 C NMR (75 MHz, CDCl 3 ) δ 165.1, 151.1, 150.6, 149.1, 142.2, 132.0, 128.3, 125.0, 121.9, 117.6, 115.7, 112.9, 111.2, 110.5, 110.1, 56.0, 56.0, 55.9, 42.9, 39.8. ESI-HRMS (m / z): calcd. for C 20 H 22 N 2 NaO 4 [M + Na] + : 377.14718; found: 377.14959.

[0092]

[0093] Example 8

[0094] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0095] The preparation methods of steps (1)-(2) are the same as those in Example 1.

[0096] (3) Synthesis of tert-butyl 6-methoxy-4-[3-(3,4,5-trimethoxyphenyl)-1-oxoprop-2-enyl]-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-7)

[0097] Using 3,4,5-trimethoxycinnamic acid (324 mg, 1.36 mmol) as the raw material, the operation process was the same as that of compound Ⅴ-1, and 343 mg of yellow solid was obtained with a yield of 62%. 1 H NMR (300 MHz, CDCl 3)δ 7.82 (d, J = 9.2 Hz, 1H), 7.67 (d, J = 15.5 Hz, 1H), 6.88 (d, J = 15.5 Hz, 1H), 6.78 (dd, J = 2.8, 9.1 Hz, 1H), 6.73 - 6.65 (m, 3H), 4.03 (t, J = 6.1 Hz, 2H), 3.87 (s, 3H), 3.87 - 3.82 (m, 8H), 3.73 (s, 3H), 1.54 (s, 9H).

[0098] (4) Synthesis of 3-(3,4,5-trimethoxyphenyl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-8)

[0099] Using Ⅴ-7 (330 mg, 0.68 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ-1, and 162 mg of yellow solid was obtained with a yield of 62%, which was the compound Ⅰ-8. 1 H NMR (300 MHz, CDCl 3 )δ 7.66 (d, J = 15.5 Hz, 1H), 6.98 (d, J = 15.5 Hz, 1H), 6.75 - 6.67 (m, 3H), 6.67 - 6.63 (m, 1H), 6.61 (d, J = 8.6 Hz, 1H), 3.98 (t, J = 4.9 Hz, 2H), 3.87 (s, 3H), 3.86 (s, 6H), 3.69 (s, 3H), 3.48 (t, J = 4.8 Hz, 2H). 13 C NMR (75 MHz, CDCl 3 )δ 164.7, 153.4, 151.1, 142.2, 139.5, 132.0, 130.9, 124.8, 119.2, 115.6, 112.9, 110.5, 105.0, 61.0, 56.1, 56.0, 42.9, 39.8. ESI-HRMS (m / z): calcd. for C 21 H 24 N 2 NaO 5 [M + Na] + : 407.15774; found: 407.15771.

[0100]

[0101] Example 9

[0102] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0103] The preparation methods of steps (1)-(2) are the same as those in Example 1.

[0104] (3) Synthesis of tert-butyl 6-methoxy-4-[3-(6-methoxypyridin-3-yl)-1-oxoprop-2-en-1-yl]-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-8)

[0105] Using 3-(6-methoxypyridin-3-yl)acrylic acid (608 mg, 3.39 mmol) as the raw material, the operation process was the same as that of compound Ⅴ-1, and 504 mg of yellow solid was obtained, with a yield of 53%. 1 H NMR(300MHz,CDCl 3 )δ8.27(s,1H),7.87 - 7.76(m,1H),7.76 - 7.66(m,2H),6.89(d,J=15.5Hz,1H),6.82 - 6.71(m,2H),6.70 - 6.62(m,1H),4.06 - 3.99(m,2H),3.96(s,3H),3.88 - 3.81(m,2H),3.74(s,3H),1.53(s,9H).

[0106] (4) Synthesis of 3-(6-methoxypyridin-3-yl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-9)

[0107] Using Ⅴ-8 (460 mg, 1.08 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ-1, and 219 mg of orange solid was obtained, with a yield of 62%, which was compound Ⅰ-9. 1 H NMR(300MHz,CDCl 3 )δ8.28(s,1H),7.77 - 7.70(m,1H),7.69 - 7.63(m,1H),6.98(d,J=15.6Hz,1H),6.73(d,J=8.7Hz,1H),6.69 - 6.63(m,2H),6.59(d,J=8.9Hz,1H),4.02 - 3.90(m,5H),3.69(s,3H),3.52 - 3.43(m,2H). 13 C NMR(75MHz,CDCl 3 )δ164.9,164.6,151.2,148.0,138.7,136.4,132.0,124.9,124.6,118.6,115.7,112.9,111.5,110.6,56.0,53.8,42.9,39.8.ESI-HRMS(m / z):calcd.for C 18 H 19 N 3 NaO3 [M+Na] + : 348.13186; found: 348.13185.

[0108]

[0109] Example 10

[0110] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0111] The preparation methods of steps (1)-(2) are the same as those in Example 1.

[0112] (3) Synthesis of tert-butyl 6-methoxy-4-[1-oxo-3-(thiophen-2-yl)prop-2-en-1-yl]-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-9)

[0113] Using 3-(thiophen-2-yl)acrylic acid (175 mg, 1.13 mmol) as the raw material, the operation process is the same as that of compound Ⅴ-1, and 209 mg of yellow solid is obtained, with a yield of 46%. 1 H NMR (300 MHz, CDCl 3 ) δ 7.88 (d, J = 15.3 Hz, 1H), 7.80 (d, J = 9.3 Hz, 1H), 7.32 (d, J = 4.9 Hz, 1H), 7.24 (d, J = 3.2 Hz, 1H), 7.07 - 7.01 (m, 1H), 6.85 - 6.75 (m, 2H), 6.71 - 6.67 (m, 1H), 4.02 (t, J = 6.2 Hz, 2H), 3.84 (t, J = 6.2 Hz, 2H), 3.75 (s, 3H), 1.53 (s, 9H).

[0114] (4) Synthesis of 1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)-3-(thiophen-2-yl)prop-2-en-1-one (Ⅰ-10)

[0115] Using Ⅴ-9 (200 mg, 0.50 mmol) as the raw material, the operation process is the same as that of the target compound Ⅰ-1, and 136 mg of yellow solid is obtained, with a yield of 91%, which is compound Ⅰ-10. 1 H NMR (300 MHz, CDCl 3)δ7.86(d,J=15.3Hz,1H),7.30(d,J=4.9Hz,1H),7.22(d,J=2.9Hz,1H),7.03(d,J=3.8,4.8Hz,1H),6.89(d,J=1 5.3Hz,1H),6.71-6.63(m,2H),6.59(d,J=9.2Hz,1H),3.96(t,J=5.0Hz,2H),3.71(s,3H),3.45(t,J=5.0Hz,2H). 13 C NMR (75MHz, CDCl 3 )δ164.6,151.3,140.6,135.0,131.9,130.3,128.1,127.6,124.7,118.6,115.8,113.5,110.1,56.1,42.9,39.8.ESI-HRMS(m / z):calcd.forC 16 H 16 N 2 NaO 2 S[M+Na] + :323.08247; found:323.08243.

[0116]

[0117] Embodiment 11

[0118] The preparation method of the α, β unsaturated ketone derivative of this embodiment comprises the following steps:

[0119] The preparation method of steps (1)-(2) is the same as that of Example 1.

[0120] (3) Synthesis of tert-butyl 4-(3-(1-(tert-butyloxycarbonyl)-1H-indol-2-yl)acryloyl)-6-methoxy-3,4-dihydroquinoxaline-1(2H)-carboxylate (V-10)

[0121] Using 3-(1-(tert-butyloxycarbonyl)-1H-indol-2-yl)acrylic acid (300 mg, 1.04 mmol) as raw material, the operation process was the same as compound V-1 to obtain 170 mg of white solid with a yield of 30%. 1 H NMR (300 MHz, CDCl 3)δ 8.11 (d, J = 8.2 Hz, 1H), 7.84 (d, J = 15.7 Hz, 1H), 7.80 - 7.71 (m, 2H), 7.63 (d, J = 7.8 Hz, 1H), 7.32 - 7.26 (m, 1H), 7.22 - 7.19 (m, 1H), 7.07 (d, J = 15.7 Hz, 1H), 6.77 - 6.71 (m, 2H), 3.99 (t, J = 6.1 Hz, 2H), 3.80 (t, J = 6.2 Hz, 2H), 3.66 (s, 3H), 1.61 (s, 9H), 1.47 (s, 9H).

[0122] (4) Synthesis of 3-(1H-indol-3-yl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-11)

[0123] Using Ⅴ-10 (180 mg, 0.34 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ-1, and 85 mg of yellow solid was obtained with a yield of 76%, which was compound Ⅰ-11. 1 H NMR (300 MHz, CDCl 3 )δ 9.18 (s, 1H), 8.11 - 7.92 (m, 1H), 7.85 - 7.68 (m, 1H), 7.50 - 7.34 (m, 2H), 7.28 - 7.13 (m, 3H), 7.00 - 6.86 (m, 1H), 6.79 - 6.51 (m, 2H), 4.02 (br, 2H), 3.69 (d, J = 8.4 Hz, 3H), 3.46 (br, 2H). 13 C NMR (75 MHz, CDCl 3 )δ 166.4, 151.2, 137.3, 136.5, 132.0, 129.0, 125.4, 125.3, 123.1, 121.3, 120.3, 115.9, 114.6, 114.0, 113.2, 112.0, 110.6, 56.1, 42.9, 40.0. ESI-HRMS (m / z): calcd. for C 20 H 19 N 3 NaO 2 [M + Na] + : 356.13695; found: 356.13696.

[0124]

[0125] Example 12

[0126] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0127] The preparation method of steps (1)-(2) is the same as that in Example 1.

[0128] (3) Synthesis of tert-butyl 6-methoxy-4-[3-(naphthalen-2-yl)-1-oxoprop-2-en-1-yl]-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-11)

[0129] Using 3-(naphthalen-2-yl)acrylic acid (200 mg, 1.01 mmol) as the raw material, the operation process is the same as that of compound Ⅴ-1, and 105 mg of yellow solid is obtained, with a yield of 23%. 1 H NMR (300 MHz, CDCl 3 ) δ 7.97 - 7.90 (m, 2H), 7.89 - 7.76 (m, 4H), 7.59 (d, J = 8.6 Hz, 1H), 7.55 - 7.47 (m, 2H), 7.11 (d, J = 15.6 Hz, 1H), 6.81 (dd, J = 2.6, 9.2 Hz, 1H), 6.77 - 6.72 (m, 1H), 4.06 (t, J = 5.9 Hz, 2H), 3.87 (t, J = 6.0 Hz, 2H), 3.73 (s, 3H), 1.54 (s, 9H).

[0130] (4) Synthesis of 3-(naphthalen-2-yl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-12)

[0131] Using Ⅴ-11 (98 mg, 0.22 mmol) as the raw material, the operation process is the same as that of the target compound Ⅰ-1, and 26 mg of yellow solid is obtained, with a yield of 34%, which is compound Ⅰ-12. 1 H NMR (300 MHz, CDCl 3 ) δ 7.97 - 7.87 (m, 2H), 7.86 - 7.74 (m, 3H), 7.59 (d, J = 8.5 Hz, 1H), 7.52 - 7.44 (m, 2H), 7.19 (d, J = 15.6 Hz, 1H), 6.77 - 6.64 (m, 2H), 6.60 (d, J = 8.6 Hz, 1H), 3.99 (t, J = 4.7 Hz, 2H), 3.67 (s, 3H), 3.47 (t, J = 5.0 Hz, 2H). 13 C NMR (75 MHz, CDCl 3)δ 164.9, 151.2, 142.4, 134.0, 133.4, 132.7, 132.0, 129.6, 128.6, 128.5, 127.8, 127.0, 126.7, 124.9, 123.6, 119.9, 115.8, 113.2, 110.3, 56.0, 42.9, 39.9. ESI-HRMS (m / z): calcd. for C 22 H 20 N 2 NaO 2 [M + Na] + : 367.14170; found: 367.14430.

[0132]

[0133] Example 13

[0134] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0135] The preparation methods of steps (1)-(2) are the same as those in Example 1.

[0136] (3) Synthesis of tert-butyl 4-[3-(benzo[d][1,3]dioxol-5-yl)-1-oxoprop-2-en-1-yl]-6-methoxy-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-12)

[0137] Using 3-(benzo[d][1,3]dioxol-5-yl)acrylic acid (200 mg, 1.04 mmol) as the raw material, the operation process is the same as that of compound Ⅴ-1, and 337 mg of yellow solid is obtained, with a yield of 74%. 1 H NMR (300 MHz, CDCl 3 ) δ 7.85 - 7.75 (m, 1H), 7.68 (d, J = 15.5 Hz, 1H), 7.02 - 6.93 (m, 2H), 6.86 - 6.74 (m, 3H), 6.70 (s, 1H), 5.99 (s, 2H), 4.02 (t, J = 5.9 Hz, 2H), 3.84 (t, J = 6.1 Hz, 2H), 3.75 (s, 3H), 1.53 (s, 9H).

[0138] (4) Synthesis of 3-(benzo[d][1,3]dioxol-5-yl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-13)

[0139] Using Ⅴ-12 (260 mg, 0.59 mmol) as the raw material, the operation process was the same as that of target compound Ⅰ-1, and 196 mg of yellow solid was obtained with a yield of 98%, namely compound Ⅰ-13. 1 H NMR (300 MHz, CDCl 3 ) δ 7.65 (d, J = 15.2 Hz, 1H), 7.07 - 6.94 (m, 2H), 6.89 (d, J = 16.0 Hz, 1H), 6.79 (d, J = 6.5 Hz, 1H), 6.72 - 6.63 (m, 2H), 6.62 - 6.54 (m, 1H), 5.98 (s, 2H), 4.04 - 3.90 (br, 2H), 3.69 (s, 3H), 3.51 - 3.41 (br, 2H). 13 C NMR (75 MHz, CDCl 3 ) δ 165.0, 151.2, 149.1, 148.3, 142.1, 132.0, 129.7, 125.0, 124.1, 117.7, 115.7, 112.9, 110.5, 108.6, 106.5, 101.5, 56.0, 42.9, 39.8. ESI-HRMS (m / z): calcd. for C 19 H 18 N 2 NaO 4 [M+Na] + : 361.11588; found: 361.11587.

[0140]

[0141] Example 14

[0142] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0143] (1) Synthesis of tert-butyl 4-benzyl-7-methoxy-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅲ-2)

[0144] Dissolve Ⅱ-2 (530 mg, 2.08 mmol), (Boc) 2 O (910 mg, 4.17 mmol), and triethylamine (269 mg, 4.17 mmol) in DCM (20 mL), and the operation process was the same as that of compound Ⅲ-1, and 603 mg of white solid was obtained with a yield of 82%. 1 H NMR (300 MHz, CDCl 3)δ 7.37 - 7.29 (m, 3H), 7.27 - 7.24 (m, 2H), 7.23 - 7.17 (m, 1H), 6.60 - 6.54 (m, 2H), 4.45 (s, 2H), 3.83 (t, J = 4.9 Hz, 2H), 3.74 (s, 3H), 3.35 (t, J = 5.0 Hz, 2H), 1.54 (s, 9H).

[0145] (2) Synthesis of tert - butyl 7 - methoxy - 3,4 - dihydroquinoxaline - 1(2H) - carboxylate (Ⅳ - 2)

[0146] Using Ⅲ - 2 (590 mg, 1.66 mmol) as the raw material, the operation process was the same as that of compound Ⅳ - 1, and 303 mg of white solid was obtained, with a yield of 72%. 1 H NMR (300 MHz, CDCl 3 )δ 7.36 - 7.30 (m, 1H), 6.68 - 6.49 (m, 2H), 3.88 - 3.70 (m, 5H), 3.41 (br, 2H), 1.58 (s, 9H).

[0147] (3) Synthesis of tert - butyl 7 - methoxy - 4 - [3 - (4 - methoxyphenyl) - 1 - oxoprop - 2 - en - 1 - yl] - 1,2,3,4 - tetrahydroquinoxaline - 1 - carboxylate (Ⅴ - 13)

[0148] Using 4 - methoxycinnamic acid (162 mg, 0.91 mmol) as the raw material, the operation process was the same as that of compound Ⅴ - 1, and 77 mg of yellow solid was obtained, with a yield of 24%. 1 H NMR (300 MHz, CDCl 3 )δ 7.63 (d, J = 15.7 Hz, 1H), 7.55 - 7.44 (m, 1H), 7.34 (d, J = 8.0 Hz, 2H), 6.97 (d, J = 8.7 Hz, 1H), 6.80 (d, J = 8.1 Hz, 2H), 6.71 (d, J = 15.4 Hz, 1H), 6.59 - 6.52 (m, 1H), 3.94 (t, J = 5.5 Hz, 2H), 3.82 - 3.71 (m, 8H), 1.48 (s, 9H).

[0149] (4) Synthesis of 3 - (4 - methoxyphenyl) - 1 - (6 - methoxy - 1,2,3,4 - tetrahydroquinoxalin - 1 - yl)prop - 2 - en - 1 - one (Ⅰ - 14)

[0150] Using Ⅴ - 13 (75 mg, 0.18 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ - 1, and 27 mg of yellow solid was obtained, with a yield of 47%, which was compound Ⅰ - 14. 1 H NMR (300 MHz, CDCl3 ) δ 7.70 (d, J = 15.6 Hz, 1H), 7.42 (d, J = 8.4 Hz, 2H), 6.98 - 6.84 (m, 4H), 6.25 (dd, J = 8.6, 2.8 Hz, 1H), 6.19 (d, J = 2.7 Hz, 1H), 3.97 (t, J = 5.2 Hz, 2H), 3.82 (s, 3H), 3.77 (s, 3H), 3.47 (t, J = 5.3 Hz, 2H). 13 C NMR (75 MHz, CDCl 3 ) δ 165.0, 160.8, 158.4, 141.6, 138.9, 129.5, 128.1, 125.8, 117.9, 117.2, 114.2, 102.2, 99.3, 55.4, 42.6, 39.2. ESI - HRMS (m / z): calcd. for C 19 H 20 N 2 O 3 [M + Na] + : 347.136621; found: 347.13662.

[0151]

[0152] Example 15

[0153] The preparation method of the α,β - unsaturated ketone derivative in this example is as follows:

[0154] The preparation methods of steps (1) and (2) are the same as those in Example 14.

[0155] (3) Synthesis of tert - butyl 7 - methoxy - 4 - [3 - (3,4,5 - trimethoxyphenyl) - 1 - oxoprop - 2 - enyl] - 1,2,3,4 - tetrahydroquinoxaline - 1 - carboxylate (Ⅴ - 14)

[0156] Using 3,4,5 - trimethoxycinnamic acid (108 mg, 0.45 mmol) as the raw material, the operation process is the same as that of compound Ⅴ - 1, and 130 mg of yellow solid is obtained, with a yield of 71%. 1 H NMR (300 MHz, CDCl 3 ) δ 7.64 - 7.51 (m, 2H), 6.95 (d, J = 8.9 Hz, 1H), 6.72 (d, J = 15.2 Hz, 1H), 6.61 (s, 2H), 6.58 - 6.51 (m, 1H), 3.95 (t, J = 6.4 Hz, 2H), 3.86 - 3.77 (m, 11H), 3.76 (s, 3H), 1.49 (s, 9H).

[0157] (4) Synthesis of 3-(3,4,5-trimethoxyphenyl)-1-(6-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)prop-2-en-1-one (Ⅰ-15)

[0158] Using Ⅴ-14 (100 mg, 0.21 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ-1, and 68 mg of yellow solid was obtained with a yield of 86%, namely compound Ⅰ-15. 1 H NMR (300 MHz, CDCl 3 ) δ 7.64 (d, J = 15.5 Hz, 1H), 6.99 - 6.85 (m, 2H), 6.70 (s, 2H), 6.28 - 6.14 (m, 2H), 4.00 - 3.94 (m, 2H), 3.89 - 3.83 (m, 9H), 3.76 (s, 3H), 3.49 (t, J = 5.2 Hz, 2H). 13 C NMR (75 MHz, CDCl 3 ) δ 164.5, 158.5, 153.4, 141.9, 139.6, 138.8, 131.0, 125.8, 119.1, 117.8, 105.1, 102.2, 99.3, 61.0, 56.2, 55.4, 42.6, 39.2. ESI-HRMS (m / z): calcd. for C 21 H 24 N 2 NaO 5 [M+Na] + : 407.15774; found: 407.15777.

[0159]

[0160] Example 16

[0161] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0162] (1) Synthesis of 1-(tert-butyl)-6-methyl-4-benzyl-3,4-dihydroquinoxaline-1,6(2H)-dicarboxylate (Ⅲ-3)

[0163] Dissolve Ⅱ-3 (450 mg, 1.59 mmol), (Boc) 2 O (695 mg, 3.19 mmol), and triethylamine (322 mg, 3.19 mmol) in DCM (20 mL), and the operation process was the same as that of compound Ⅲ-1, and 475 mg of yellow solid was obtained with a yield of 78%. 1 H NMR (300 MHz, CDCl 3) δ 7.59 - 7.54 (m, 1H), 7.40 - 7.39 (m, 1H), 7.37 - 7.31 (m, 3H), 7.30 - 7.23 (m, 3H), 4.57 (s, 2H), 3.87 - 3.81 (m, 5H), 3.41 (t, J = 5.0 Hz, 2H), 1.53 (s, 9H).

[0164] (2) Synthesis of 1-(tert-Butyl)-6-methyl-3,4-dihydroquinoxaline-1,6(2H)-dicarboxylate (IV-3)

[0165] Using III-3 (460 mg, 1.20 mmol) as the raw material, the operation process was the same as that of compound IV-1, and 330 mg of colorless oil was obtained with a yield of 94%. 1 H NMR (300 MHz, CDCl 3 ) δ 7.59 (d, J = 8.4 Hz, 1H), 7.32 (dd, J = 1.8, 8.6 Hz, 1H), 7.27 - 7.24 (m, 1H), 3.87 (s, 3H), 3.79 (t, J = 4.8 Hz, 2H), 3.42 (t, J = 5.0 Hz, 2H), 1.53 (s, 9H).

[0166] (3) Synthesis of Methyl 4-[3-(4-methoxyphenyl)-1-oxoprop-2-enyl]-1-{[(2-methylpropan-2-yl)oxy]carbonyl}-1,2,3,4-tetrahydroquinoxaline-6-carboxylate (V-15)

[0167] Using 4-methoxycinnamic acid (61 mg, 0.34 mmol) as the raw material, the operation process was the same as that of compound V-1, and 100 mg of yellow solid was obtained with a yield of 65%. 1 H NMR (300 MHz, CDCl 3 ) δ 8.04 (d, J = 8.6 Hz, 1H), 7.84 - 7.77 (m, 2H), 7.71 (d, J = 15.5 Hz, 1H), 7.34 (d, J = 8.7 Hz, 2H), 6.80 (d, J = 8.7 Hz, 2H), 6.70 (d, J = 15.5 Hz, 1H), 3.97 (t, J = 5.6 Hz, 2H), 3.86 - 3.81 (m, 2H), 3.80 (s, 3H), 3.75 (s, 3H), 1.48 (s, 9H).

[0168] (4) Synthesis of Methyl 4-[3-(4-methoxyphenyl)-1-oxoprop-2-enyl]-1,2,3,4-tetrahydroquinoxaline-6-carboxylate (I-16)

[0169] Using Ⅴ-15 (100 mg, 0.22 mmol) as the raw material, following the same operation process as that for the target compound Ⅰ-1, 40 mg of yellow solid was obtained with a yield of 51%, namely compound Ⅰ-16. 1 H NMR (300 MHz, CDCl 3 ) δ 7.85 - 7.72 (m, 3H), 7.45 (d, J = 8.6 Hz, 2H), 6.95 - 6.87 (m, 3H), 6.65 (d, J = 8.5 Hz, 1H), 4.03 (t, J = 4.7 Hz, 2H), 3.85 (s, 6H), 3.57 (t, J = 5.0 Hz, 2H). 13 C NMR (75 MHz, CDCl 3 ) δ 166.9, 165.1, 161.0, 142.8, 141.9, 129.7, 128.3, 128.0, 126.6, 123.1, 117.6, 116.3, 114.3, 113.4, 55.4, 51.8, 42.5, 38.4. ESI-HRMS (m / z): calcd. for C 20 H 20 N 2 NaO 4 [M + Na] + : 375.13153; found: 375.13152.

[0170]

[0171] Example 17

[0172] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0173] The preparation methods of steps (1) and (2) are the same as those in Example 16.

[0174] (3) Synthesis of methyl 4-[3-(3,4,5-trimethoxyphenyl)-1-oxoprop-2-enyl]-1-{[(2-methylpropan-2-yl)oxy]carbonyl}-1,2,3,4-tetrahydroquinoxaline-6-carboxylate (Ⅴ-16)

[0175] Using 3,4,5-trimethoxycinnamic acid (489 mg, 2.05 mmol) as the raw material, following the same operation process as that for compound Ⅴ-1, 560 mg of yellow solid was obtained with a yield of 80%. 1 H NMR (300 MHz, CDCl 3)δ8.12(d, J = 8.7Hz, 1H), 7.91 - 7.85(m, 2H), 7.73(d, J = 15.4Hz, 1H), 6.82(d, J = 15.4Hz, 1H), 6.69(s, 2H), 4.06(t, J = 5.9Hz, 2H), 3.94 - 3.90(m, 2H), 3.87(s, 3H), 3.86(s, 3H), 3.84(s, 6H), 1.56(s, 9H).

[0176] (4) Synthesis of Methyl 4 - [3 - (3,4,5 - Trimethoxyphenyl) - 1 - oxoprop - 2 - enyl] - 1,2,3,4 - tetrahydroquinoxaline - 6 - carboxylate (Ⅰ - 17)

[0177] Using Ⅴ - 16 (330 mg, 0.64 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ - 1, and 238 mg of yellow solid was obtained with a yield of 90%, which was compound Ⅰ - 17. 1 H NMR (300 MHz, CDCl 3 )δ7.81(s, 1H), 7.75 - 7.71(m, 1H), 7.71 - 7.67(m, 1H), 6.95(d, J = 15.4Hz, 1H), 6.71(s, 2H), 6.63(d, J = 8.5Hz, 1H), 4.01(t, J = 5.1Hz, 2H), 3.87(s, 3H), 3.84(s, 6H), 3.81(s, 3H), 3.54(t, J = 5.2Hz, 2H). 13 C NMR (75 MHz, CDCl 3 )δ166.8, 164.6, 153.4, 142.9, 142.0, 139.5, 130.9, 128.3, 126.6, 122.9, 118.4, 117.3, 113.4, 105.1, 61.0, 56.1, 51.8, 42.4, 38.3. ESI - HRMS (m / z): calcd. for C 22 H 24 N 2 NaO 6 [M + Na] + : 435.15266; found: 435.15266.

[0178]

[0179] Example 18

[0180] The preparation method of the α,β - unsaturated ketone derivative in this example is as follows:

[0181] (1) Synthesis of 1-(tert-Butyl)-7-methyl-4-benzyl-3,4-dihydroquinoxaline-1,7(2H)-dicarboxylate (Ⅲ-4)

[0182] Dissolve Ⅱ-4 (957 mg, 3.39 mmol), (Boc) 2 O (1.48 g, 6.78 mmol), DIPEA (876 mg, 6.78 mmol), and DMAP (83 mg, 0.68 mmol) in DCM (30 mL). The operation process is the same as that of compound Ⅲ-1 to obtain 1.06 g of yellow solid with a yield of 82%. 1 H NMR (300 MHz, CDCl 3 ) δ 8.15 (s, 1H), 7.61 (dd, J = 1.4, 8.7 Hz, 1H), 7.38 - 7.25 (m, 3H), 7.24 - 7.17 (m, 2H), 6.60 (d, J = 8.7 Hz, 1H), 4.59 (s, 2H), 3.90 - 3.77 (m, 5H), 3.51 (t, J = 5.0 Hz, 2H), 1.55 (s, 9H).

[0183] (2) Synthesis of 1-(tert-Butyl)-7-methyl-3,4-dihydroquinoxaline-1,7(2H)-dicarboxylate (Ⅳ-4)

[0184] Using Ⅲ-4 (1.00 g, 2.61 mmol) as the raw material, the operation process is the same as that of compound Ⅳ-1 to obtain 730 mg of white solid with a yield of 96%. 1 H NMR (300 MHz, CDCl 3 ) δ 8.18 (s, 1H), 7.60 (dd, J = 1.9, 8.5 Hz, 1H), 6.52 (d, J = 8.5 Hz, 1H), 4.46 (s, 1H), 3.85 (s, 3H), 3.77 (t, J = 4.7 Hz, 2H), 3.46 (t, J = 5.4 Hz, 2H), 1.54 (s, 9H).

[0185] (3) Synthesis of tert-Butyl 7-(methoxycarbonyl)-4-[3-(4-methoxyphenyl)-1-oxoprop-2-en-1-yl]-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-17)

[0186] Using 4-methoxycinnamic acid (183 mg, 1.03 mmol) as the raw material, the operation process is the same as that of compound Ⅴ-1 to obtain 207 mg of yellow solid with a yield of 67%. 1 H NMR (300 MHz, CDCl 3)δ8.62(s,1H),7.81 - 7.71(m,2H),7.42(d,J=8.6Hz,2H),7.24(d,J=14.9Hz,1H),6.88(d,J=8.6Hz,2H),6.76(d,J=15.5Hz,1H),4.06(t,J=5.8Hz,2H),3.96 - 3.87(m,5H),3.83(s,3H),1.57(s,9H).

[0187] (4) Synthesis of Methyl 1 - [3 - (4 - methoxyphenyl) - 1 - oxoprop - 2 - enyl] - 1,2,3,4 - tetrahydroquinoxaline - 6 - carboxylate (Ⅰ - 18)

[0188] Using Ⅴ - 17 (200 mg, 0.44 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ - 1, and 97 mg of yellow solid was obtained with a yield of 62%, which was compound Ⅰ - 18. 1 H NMR(300MHz,CDCl 3 )δ7.73(d,J=15.5Hz,1H),7.42(d,J=8.5Hz,2H),7.37 - 7.30(m,2H),7.14 - 7.06(m,1H),6.93 - 6.80(m,3H),4.33(s,1H),3.99(t,J=4.8Hz,2H),3.90(s,3H),3.83(s,3H),3.53(t,J=4.7Hz,2H). 13 C NMR(75MHz,CDCl 3 )δ166.9,165.1,161.1,142.6,137.4,129.6,128.1,127.8,127.5,124.4,117.5,116.7,115.5,114.3,55.4,52.1,42.4,38.8.ESI - HRMS(m / z):calcd.for C 20 H 20 N 2 NaO 4 [M + Na] + :375.13153;found:375.13158.

[0189]

[0190] Example 19

[0191] The preparation method of the α,β - unsaturated ketone derivative in this example is as follows:

[0192] The preparation methods of steps (1) and (2) are the same as those in Example 18.

[0193] (3)Synthesis of tert-butyl 4-[2-cyano-1-oxo-3-(3,4,5-trimethoxyphenyl)prop-2-enyl]-7-(methoxycarbonyl)-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-18)

[0194] Using 3,4,5-trimethoxycinnamic acid (330 mg, 1.37 mmol) as the raw material, the operation process was the same as that of compound Ⅴ-1 to obtain 345 mg of yellow solid with a yield of 39%. 1 H NMR(300MHz,CDCl 3 )δ8.63(s,1H),7.76-7.67(m,2H),7.25(d,J=15.8Hz,1H),6.77(d,J=15.5Hz,1H),6.69(s,2H),4.06(t,J=5.9Hz,2H),3.94-3.90(m,5H),3.88(s,3H),3.86(s,6H),1.58(s,9H).

[0195] (4)Synthesis of methyl 1-[3-(3,4,5-methoxyphenyl)-1-oxoprop-2-enyl]-1,2,3,4-tetrahydroquinoxaline-6-carboxylate (Ⅰ-19)

[0196] Using Ⅴ-18 (325 mg, 0.63 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ-1 to obtain 217 mg of yellow solid with a yield of 83%, which was compound Ⅰ-19. 1 H NMR(300MHz,CDCl 3 )δ7.69(d,J=15.4Hz,1H),7.38-7.26(m,2H),7.09(d,J=8.0Hz,1H),6.86(d,J=15.5Hz,1H),6.70(s,2H),4.38(s,1H),4.05-3.96(m,2H),3.89(s,3H),3.87(s,3H),3.86(s,6H),3.59-3.51(m,2H). 13 C NMR(75MHz,CDCl 3 )δ166.9,164.7,153.4,143.0,139.8,137.4,130.6,127.9,127.7,124.3,118.6,117.4,115.6,105.2,61.0,56.2,52.1,42.5,38.9.ESI-HRMS(m / z):calcd.forC 22 H 24 N 2 NaO 6 [M+Na]+ : 435.15266; found: 435.15269.

[0197]

[0198] Example 20

[0199] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0200] The preparation methods of steps (1) and (2) are the same as those in Example 18.

[0201] (3) Synthesis of tert-butyl 4-[3-(3-hydroxy-4-methoxyphenyl)-1-oxoprop-2-enyl]-7-(methoxycarbonyl)-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-19)

[0202] Using 3-hydroxy-4-methoxycinnamic acid (200 mg, 1.03 mmol) as the raw material, the operation process is the same as that of compound Ⅴ-1, and 215 mg of yellow solid is obtained, with a yield of 67%. 1 H NMR (300 MHz, CDCl 3 ) δ 8.61 (d, J = 1.3 Hz, 1H), 7.79 - 7.65 (m, 2H), 7.22 (d, J = 8.4 Hz, 1H), 7.05 (d, J = 1.9 Hz, 1H), 7.00 (dd, J = 2.0, 8.3 Hz, 1H), 6.83 (d, J = 8.3 Hz, 1H), 6.74 (d, J = 15.4 Hz, 1H), 5.70 (s, 1H), 4.05 (t, J = 5.9 Hz, 2H), 3.93 (s, 3H), 3.91 (s, 3H), 3.90 - 3.86 (m, 2H), 1.57 (s, 9H).

[0203] (4) Synthesis of methyl 1-[3-(3-hydroxy-4-methoxyphenyl)-1-oxoprop-2-enyl]-1,2,3,4-tetrahydroquinoxaline-6-carboxylate (Ⅰ-20)

[0204] Using Ⅴ-19 (215 mg, 0.46 mmol) as the raw material, the operation process is the same as that of the target compound Ⅰ-1, and 38 mg of yellow solid is obtained, with a yield of 23%, which is the compound Ⅰ-20. 1 H NMR (300 MHz, CDCl 3 +D 2O) δ 7.68 (d, J = 15.5 Hz, 1H), 7.37 - 7.31 (m, 2H), 7.09 (d, J = 8.8 Hz, 1H), 7.06 (d, J = 1.8 Hz, 1H), 7.00 (dd, J = 1.7, 8.4 Hz, 1H), 6.84 (s, 1H), 6.80 (d, J = 6.4 Hz, 1H), 3.99 (t, J = 5.0 Hz, 2H), 3.92 (s, 3H), 3.90 (s, 3H), 3.52 (t, J = 5.1 Hz, 2H). 13 C NMR (75 MHz, DMSO-d 6 ) δ 166.7, 164.4, 150.1, 147.2, 142.6, 138.9, 128.1, 127.6, 126.9, 124.7, 121.6, 117.0, 115.9, 115.0, 113.8, 112.4, 56.0, 52.4, 41.8. ESI-HRMS (m / z): calcd. for C 20 H 20 N 2 NaO 5 [M + Na] + : 391.12644; found: 391.12642.

[0205]

[0206] Example 21

[0207] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0208] The preparation methods of steps (1) and (2) are the same as those in Example 18.

[0209] (3) Synthesis of tert-butyl 4-[3-(4-hydroxy-3-methoxyphenyl)-1-oxoprop-2-enyl]-7-(methoxycarbonyl)-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-20)

[0210] Using 4-hydroxy-3-methoxycinnamic acid (219 mg, 1.13 mmol) as the raw material, the operation process is the same as that of compound Ⅴ-1, and 261 mg of yellow solid is obtained with a yield of 54%. 1 H NMR (300 MHz, CDCl 3)δ8.62(d,J=1.3Hz,1H),7.77-7.74(m,1H),7.73-7.69(m,1H),7.23(d,J=8.3Hz,1H),7.04(dd,J=1.6,8.3Hz,1H),6.94(d,J=1.5Hz,1H),6.90(d,J=8.2Hz,1H),6.72(d,J=15.5Hz,1H),5.85(s,1H),4.05(t,J=5.9Hz,2H),3.95-3.86(m,8H),1.57(s,9H).

[0211] (4) Synthesis of methyl 1-[3-(4-hydroxy-3-methoxyphenyl)-1-oxoprop-2-enyl]-1,2,3,4-tetrahydroquinoxaline-6-carboxylate (Ⅰ-21)

[0212] Using Ⅴ-20 (250 mg, 0.53 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ-1, and 156 mg of yellow solid was obtained with a yield of 79%, which was compound Ⅰ-21. 1 H NMR(300MHz,CDCl 3 )δ7.70(d,J=15.4Hz,1H),7.44-7.29(m,2H),7.17-7.01(m,2H),6.99-6.87(m,2H),6.81(d,J=15.5Hz,1H),6.00(s,1H),4.34(s,1H),4.07-3.96(m,2H),3.89(s,3H),3.88(s,3H),3.60-3.45(m,2H). 13 C NMR(75MHz,CDCl 3 )δ167.0,165.1,147.7,146.8,143.2,137.5,128.1,127.6,124.4,122.3,117.5,116.7,115.6,114.9,110.1,56.0,52.1,42.5,38.9.ESI-HRMS(m / z):calcd.forC 20 H 20 N 2 NaO 5 [M+Na] + :391.12644;found:391.12655.

[0213]

[0214] Example 22

[0215] The preparation method of the α,β-unsaturated ketone derivative of this example is as follows:

[0216] The preparation methods of steps (1) and (2) are the same as those in Example 18.

[0217] (3) Synthesis of tert-butyl 7-(methoxycarbonyl)-4-(1-oxoprop-2-en-1-yl)-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-21)

[0218] Dissolve Ⅳ-4 (100 mg, 0.34 mmol) and DIPEA (66 mg, 0.51 mmol) in anhydrous DCM (4 mL) solution, and dropwise add anhydrous DCM solution of acryloyl chloride (33 mg, 0.36 mmol) at 0 °C, and stir at room temperature for 12 h. Monitor by TLC (DCM:MeOH = 50:1). After the reaction is complete, dilute with DCM (20 mL), wash with 1N HCl (20 mL × 3), water (20 mL × 3), 1N NaOH (20 mL × 3) and saturated NaCl (20 mL × 3), and dry over anhydrous Na 2 SO 4 for overnight. Filter and concentrate to obtain the crude product, and purify by column chromatography (PE:EtOAc = 6:1) to obtain 60 mg of yellow solid, with a yield of 51%. 1 1H NMR (300 MHz, CDCl 3 ) δ 8.62 (s, 1H), 7.77 (dd, J = 1.5, 8.4 Hz, 1H), 7.19 (d, J = 8.3 Hz, 1H), 6.65 (dd, J = 9.4, 16.7 Hz, 1H), 6.55 (dd, J = 2.5, 16.8 Hz, 1H), 5.83 (dd, J = 2.5, 9.5 Hz, 1H), 4.06 (t, J = 5.9 Hz, 2H), 3.96 (s, 3H), 3.91 (t, J = 6.1 Hz, 2H), 1.60 (s, 9H).

[0219] (4) Synthesis of methyl 1-(1-oxoprop-2-en-1-yl)-1,2,3,4-tetrahydroquinoxaline-6-carboxylate (Ⅰ-22)

[0220] Using Ⅴ-21 (50 mg, 0.14 mmol) as the raw material, the operation process is the same as that of the target compound Ⅰ-1, to obtain 20 mg of yellow solid, with a yield of 56%, that is, compound Ⅰ-22. 1 1H NMR (300 MHz, CDCl 3) δ 7.34 - 7.28 (m, 2H), 7.02 (d, J = 8.1 Hz, 1H), 6.67 (dd, J = 10.1, 16.8 Hz, 1H), 6.48 (dd, J = 1.9, 16.8 Hz, 1H), 5.76 (dd, J = 1.9, 10.1 Hz, 1H), 4.26 (s, 1H), 3.95 (t, J = 5.0 Hz, 2H), 3.89 (s, 3H), 3.55 - 3.48 (m, 2H). 13 C NMR (75 MHz, CDCl 3 ) δ 166.8, 164.4, 137.3, 129.2, 128.7, 127.8, 127.5, 124.4, 117.5, 115.5, 52.1, 42.3, 38.8. ESI - HRMS (m / z): calcd. for C 13 H 14 N 2 NaO 3 [M + Na] + : 269.08966; found: 269.08968.

[0221]

[0222] Example 23

[0223] The preparation method of the α,β - unsaturated ketone derivative in this example is as follows:

[0224] The preparation methods of steps (1) - (4) are the same as those in Example 19.

[0225] (5) Synthesis of 1 - [3 - (3,4,5 - trimethoxyphenyl) - 1 - oxoprop - 2 - enyl] - 1,2,3,4 - tetrahydroquinoxaline - 6 - carboxamide (Ⅰ - 23)

[0226] Dissolve Ⅰ - 19 (100 mg, 0.24 mmol) and LiOH (9 mg, 0.36 mmol) in 2 mL of MeOH:H 2 O (3:1) and reflux for 5 h. Cool to room temperature, rotary evaporate MeOH, add 1 mol / L HCl to adjust the pH to 3, and filter. The filter cake is dried to obtain 76 mg of yellow solid, with a yield of 78%, without further purification. 1 H NMR (300 MHz, DMSO - d 6) δ 12.61 (broad, 1H), 7.56 (doublet, J = 15.2 Hz, 1H), 7.28 (singlet, 1H), 7.22 - 7.14 (multiplet, 1H), 7.13 - 7.01 (multiplet, 2H), 6.97 (singlet, 2H), 6.53 (singlet, 1H), 3.93 - 3.84 (multiplet, 2H), 3.80 (singlet, 6H), 3.69 (singlet, 3H).

[0227] Dissolve the obtained yellow solid (70 mg, 0.18 mmol) and CDI (34 mg, 0.2 mmol) in anhydrous THF (2 mL), and reflux overnight under nitrogen protection. Cool the mixture to room temperature, add ammonium hydroxide (1 mL), stir at room temperature for 12 h, remove the solvent, dissolve in DCM, wash with water (10 mL × 3) and saturated NaCl (10 mL × 3), and dry over anhydrous Na 2 SO 4 Dry, filter by suction, and concentrate. Purify by column chromatography (DCM:MeOH = 500:1) to obtain 35 mg of yellow solid with a yield of 50%, which is compound I - 23. 1 1H NMR (300 MHz, CDCl 3 ) δ 7.69 (doublet, J = 15.6 Hz, 1H), 7.19 (singlet, 1H), 7.09 (doublet, J = 8.3 Hz, 1H), 7.00 (doublet, J = 8.3 Hz, 1H), 6.84 (doublet, J = 15.4 Hz, 1H), 6.70 (singlet, 2H), 6.12 (broad, 1H), 5.86 (broad, 1H), 4.43 (broad, 1H), 3.99 (triplet, J = 5.1 Hz, 2H), 3.87 (singlet, 3H), 3.86 (singlet, 6H), 3.54 (triplet, J = 5.0 Hz, 2H). 13 13C NMR (75 MHz, CDCl 3 ) δ 169.1, 164.8, 153.5, 143.1, 139.9, 137.9, 131.1, 130.7, 127.1, 124.4, 118.6, 114.6, 114.0, 105.3, 61.0, 56.3, 42.5, 29.7. ESI - HRMS (m / z): calculated for C 21 H 23 N 3 NaO 5 [M + Na] + : 420.15299; found: 420.15300.

[0228]

[0229] Example 24

[0230] The preparation method of the α,β-unsaturated ketone derivative of this embodiment is as follows:

[0231] The preparation methods of steps (1) and (2) are the same as those in Embodiment 1.

[0232] (3) Synthesis of tert-butyl 6-methoxy-4-[3-(4-methoxyphenyl)-2-methyl-1-oxoprop-2-en-1-yl]-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-22)

[0233] Using 3-(4-methoxyphenyl)-2-methylacrylic acid (436 mg, 2.27 mmol) as the raw material, the operation process is the same as that of compound Ⅴ-1, and 402 mg of yellow solid is obtained, with a yield of 61%. 1 H NMR(300MHz,CDCl 3 )δ7.69(d,J=7.5Hz,1H),7.21(d,J=8.7Hz,2H),6.88(d,J=8.7Hz,2H),6.80(s,1H),6.73(d,J=2.8Hz,1H),6.68(dd,J=2.8,9.0Hz,1H),3.98(t,J=5.9Hz,2H),3.87(t,J=6.0Hz,2H),3.81(s,3H),3.65(s,3H),1.96(d,J=0.9Hz,3H),1.52(s,9H).

[0234] (4) Synthesis of 3-(4-methoxyphenyl)-1-(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)-2-methylprop-2-en-1-one (Ⅰ-24)

[0235] Using Ⅴ-22 (386 mg, 0.88 mmol) as the raw material, the operation process is the same as that of the target compound Ⅰ-1, and 182 mg of yellow oil is obtained, with a yield of 61%, which is compound Ⅰ-24. 1 H NMR(300MHz,CDCl 3 )δ7.24(d,J=8.5Hz,2H),6.87(d,J=8.6Hz,2H),6.85-6.75(m,2H),6.60-6.49(m,2H),3.90(t,J=4.6Hz,2H),3.79(s,3H),3.60(s,3H),3.43(t,J=4.9Hz,2H),2.02(s,3H). 13 C NMR(75MHz,CDCl 3)δ170.8,158.0,150.2,132.6,130.6,130.1,129.5,127.6,124.4,114.8,112.8,111.8,108.2,54.8,54.2,41.7,40.5,15.2.ESI-HRMS(m / z):calcd.for C 20 H 22 N 2 NaO 3 [M+Na] + :361.15226;found:361.15225.

[0236]

[0237] Example 25

[0238] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0239] The preparation methods of steps (1) and (2) are the same as those in Example 1.

[0240] (3) Synthesis of tert-butyl 4-[2-cyano-3-(4-methoxyphenyl)-1-oxoprop-2-enyl]-6-methoxy-1,2,3,4-tetrahydroquinoxaline-1-carboxylate (Ⅴ-23)

[0241] Using 2-cyano-3-(4-methoxyphenyl)acrylic acid (138 mg, 0.68 mmol) as the raw material, the operation process is the same as that of Compound Ⅴ-1, and 145 mg of yellow solid is obtained, with a yield of 57%. 1 H NMR(300MHz,CDCl 3 )δ7.95 - 7.84(m,3H),7.81 - 7.68(m,1H),6.96(d,J=8.3Hz,2H),6.81 - 6.73(m,1H),6.70(s,1H),4.08 - 3.99(m,2H),3.95 - 3.85(m,5H),3.70(s,3H),1.52(s,9H).

[0242] (4) Synthesis of 3-(4-methoxyphenyl)-2-[(7-methoxy-1,2,3,4-tetrahydroquinoxalin-1-yl)carbonyl]prop-2-enenitrile (Ⅰ-25)

[0243] Using Ⅴ-23 (120 mg, 0.27 mmol) as the raw material, the operation process is the same as that of the target compound Ⅰ-1, and 80 mg of yellow oil is obtained, with a yield of 86%, which is Compound Ⅰ-25. 1 H NMR(300MHz,CDCl 3)δ 7.97 - 7.84 (m, 3H), 6.96 (d, J = 7.7 Hz, 2H), 6.75 - 6.63 (m, 2H), 6.60 (d, J = 8.5 Hz, 1H), 3.97 (br, 2H), 3.88 (s, 3H), 3.64 (s, 3H), 3.52 (br, 2H). 13 C NMR (75 MHz, CDCl 3 )δ 163.1, 162.5, 152.7, 151.6, 132.7, 131.4, 125.1, 124.4, 116.2, 115.9, 114.7, 113.7, 109.7, 104.5, 56.0, 55.6, 42.6, 42.5. ESI - HRMS (m / z): calcd. for C 20 H 19 N 3 NaO 3 [M + Na] + : 372.13186; found: 372.13151.

[0244]

[0245] Example 26

[0246] The preparation method of the α,β - unsaturated ketone derivative in this example is as follows:

[0247] The preparation methods of steps (1) and (2) are the same as those in Example 1.

[0248] (3) Synthesis of tert - butyl 4 - [2 - cyano - 3 - (4 - methoxyphenyl) - 1 - oxoprop - 2 - enyl] - 7 - (methoxycarbonyl) - 1,2,3,4 - tetrahydroquinoxaline - 1 - carboxylate (Ⅴ - 24)

[0249] Using 2 - cyano - 3 - (4 - methoxyphenyl)acrylic acid (300 mg, 1.48 mmol) as the raw material, the operation process is the same as that of compound Ⅴ - 1, and 383 mg of yellow solid is obtained with a yield of 54%. 1 H NMR (300 MHz, CDCl 3 )δ 8.63 (s, 1H), 7.96 (s, 1H), 7.91 (d, J = 6.8 Hz, 2H), 7.70 (d, J = 6.5 Hz, 1H), 7.31 - 7.17 (m, 1H), 6.97 (d, J = 6.1 Hz, 2H), 4.05 (br, 2H), 3.96 (br, 2H), 3.90 (s, 3H), 3.89 (s, 3H), 1.57 (s, 9H).

[0250] (4) Synthesis of methyl 1-[2-cyano-3-(4-methoxyphenyl)-1-oxoprop-2-enyl]-1,2,3,4-tetrahydroquinoxaline-6-carboxylate (Ⅰ-26)

[0251] Using Ⅴ-24 (200 mg, 0.42 mmol) as the raw material, the operation process was the same as that of the target compound Ⅰ-1, and 137 mg of yellow solid was obtained with a yield of 87%, which was the compound Ⅰ-26. 1 H NMR (300 MHz, CDCl 3 ) δ 7.94 (s, 1H), 7.90 (d, J = 8.4 Hz, 2H), 7.34 (s, 1H), 7.29 (d, J = 9.0 Hz, 1H), 7.10 (d, J = 8.1 Hz, 1H), 6.96 (d, J = 8.3 Hz, 2H), 4.39 (br, 1H), 4.03 - 3.94 (m, 2H), 3.87 (s, 3H), 3.86 (s, 3H), 3.63 - 3.52 (m, 2H). 13 C NMR (75 MHz, CDCl 3 ) δ 166.8, 163.3, 162.7, 153.3, 136.9, 132.9, 128.2, 127.4, 125.0, 123.4, 118.1, 116.1, 115.9, 114.7, 103.9, 55.6, 52.1, 42.2, 41.4. ESI-HRMS (m / z): calcd. for C 21 H 19 N 3 NaO 4 [M + Na] + : 400.12678; found: 400.12691.

[0252]

[0253] Example 27

[0254] The preparation method of the α,β-unsaturated ketone derivative in this example is as follows:

[0255] (1) Synthesis of 4-{[(2-methylpropan-2-yl)oxy]carbonyl}-1,2,3,4-tetrahydroquinoxaline-6-carboxylic acid (Ⅵ-1)

[0256] Dissolve Ⅳ-4 (45 mg, 0.15 mmol) and LiOH (12 mg, 0.46 mmol) in 2 mL of MeOH:H 2In O(3:1), reflux and stir. Monitor the reaction by TLC (PE:EtOAc = 1:1). After 15 h, the reaction is complete, stop the reaction. Cool to room temperature, add 1N HCl dropwise to adjust the pH to about 3, and white solid precipitates. Filter by suction, dry the filter cake to obtain 32 mg of white solid, with a yield of 74.8%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 12.05 (br, 1H), 7.96 (s, 1H), 7.41 (d, J = 8.7 Hz, 1H), 6.89 (s, 1H), 6.57 (d, J = 8.4 Hz, 1H), 3.65 - 3.56 (m, 2H), 3.31 - 3.27 (m, 2H), 1.46 (s, 9H).

[0257] (2) Synthesis of ethyl 4-{[(2-methylpropan-2-yl)oxy]carbonyl}-1,2,3,4-tetrahydroquinoxaline-6-carboxylate (Ⅶ-1)

[0258] Dissolve Ⅵ-1 (30 mg, 0.11 mmol) in EtOH, stir at room temperature for 10 min, then add SOCl 2 solution (26 mg, 0.22 mmol) dropwise at 0 °C. Monitor the reaction by TLC (PE:EtOAc = 5:1). After 20 h, the reaction is complete, stop the reaction. Rotavapor to dryness, add 10 mL of EtOAc to dissolve, wash with saturated NaHCO 3 (10 mL × 3), water (10 mL × 3), saturated NaCl (10 mL × 3), and dry over anhydrous Na 2 SO 4 overnight. Filter by suction, rotary evaporate the filtrate to dryness, and white solid precipitates in an ice bath, 29 mg, with a yield of 87.9%. 1 H NMR (300 MHz, CDCl 3 ) δ 8.16 (s, 1H), 7.60 (dd, J = 8.6, 1.4 Hz, 1H), 6.52 (d, J = 8.4 Hz, 1H), 4.45 (br, 1H), 4.31 (q, J = 7.1 Hz, 2H), 3.77 (t, J = 4.7 Hz, 2H), 3.50 - 3.42 (m, 2H), 1.54 (s, 9H), 1.36 (t, J = 7.1 Hz, 3H).

[0259] (3) Synthesis of ethyl 1-[3-(3-hydroxy-4-methoxyphenyl)-1-oxoprop-2-enyl]-4-{[(2-methylpropan-2-yl)oxy]carbonyl}-1,2,3,4-tetrahydroquinoxaline-6-carboxylate (Ⅴ-25)

[0260] Using 3-hydroxy-4-methoxycinnamic acid (399 mg, 2.05 mmol) as the raw material, the operation process was the same as that of compound V-1. After treatment, the reaction solution was concentrated to dryness without purification and directly used in the next step.

[0261] (4) Synthesis of ethyl 1-[3-(3-hydroxy-4-methoxyphenyl)-1-oxoprop-2-enyl]-1,2,3,4-tetrahydroquinoxaline-6-carboxylate (I-27)

[0262] Using V-25 (unpurified) as the raw material, the operation process was the same as that of the target compound I-1 to obtain 48 mg of yellow solid. The yield of the two-step reaction was 7%, namely compound I-27. 1 H NMR (300 MHz, CDCl 3 ) δ 7.68 (d, J = 15.5 Hz, 1H), 7.37 - 7.31 (m, 2H), 7.11 - 7.05 (m, 2H), 7.00 (d, J = 8.2 Hz, 1H), 6.84 (d, J = 2.2 Hz, 1H), 6.80 (d, J = 4.4 Hz, 1H), 5.79 (s, 1H), 4.36 (q, J = 7.1 Hz, 2H), 3.99 (t, J = 4.9 Hz, 2H), 3.91 (s, 3H), 3.52 (t, J = 4.9 Hz, 2H), 1.39 (t, J = 7.1 Hz, 3H). 13 C NMR (75 MHz, CDCl 3 ) δ 166.5, 165.1, 148.3, 145.8, 142.8, 137.4, 128.7, 128.0, 127.9, 124.3, 121.8, 117.6, 117.3, 115.6, 112.9, 110.6, 61.0, 56.0, 42.5, 38.9, 14.4. ESI-HRMS (m / z): calcd. for C 21 H 22 O 5 N 2 [M+Na] + : 405.14209; found: 405.14016.

[0263]

[0264] Example of implementation effect

[0265] I. Proliferation inhibition experiment on tumor cells

[0266] Test the proliferation inhibitory activity of the compounds of the present invention against human fibrosarcoma cells (HT-1080), human breast cancer cells (MDA-MB-231, MCF-7), and human cervical cancer cells (HeLa). All of the above cell lines were cryopreserved and passaged in our laboratory. Tumor cells in the logarithmic growth phase were seeded into 96-well plates at 2000 - 4000 cells / well. After 24 h, different concentrations of the target compound were added, and the cells were cultured at 37°C and 5% CO 2 for 48 h. 50 μL of MTT solution (5 mg / mL) was added to each well, and after continued culturing for 4 h, the culture medium and MTT solution were discarded. 100 μL of DMSO solution was added to each well. After shaking, the absorbance (OD value) was measured at a wavelength of 570 nm using an enzyme-linked immunosorbent assay (ELISA) reader. Calculate the cell inhibition rate: Growth inhibition rate = (OD control - OD experiment) / (OD control - OD blank) × 100% (OD control, OD experiment, and OD blank represent the average light absorption values of each group), and then use GraphPad software to fit the IC 50 of the compound. The experimental results are shown in Table 1, indicating that some compounds have strong proliferation inhibitory activity against tumor cells.

[0267] Table 1 Proliferation inhibitory activity of some compounds against three tumor cell lines

[0268]

[0269]

[0270] II. Cell cycle experiment

[0271] Test the effect of the compounds of the present invention on the cell cycle of cancer cells (HT-1080). HT-1080 cells were seeded onto 6-well plates at an appropriate concentration and incubated at 37°C and 5% CO 2 for 24 h. Then, different concentrations of the compound or positive control were added to each well, and the cells were continued to be cultured at 37°C and 5% CO 2 for another 24 h. The cells were collected, centrifuged, and slowly and vertically dropped into 10 mL of pre-cooled 70% (v / v) ethanol for fixation at -20°C for 24 h. After washing with PBS, 500 μL of staining agent (50 μg / mL of PI and 100 μg / mL of RNase A) was added to each sample, and the samples were incubated at 4°C in the dark for 1 h. After centrifugation, the dye was removed, the samples were washed with PBS, filtered through a 300-mesh filter, and detected at 580 nm using a flow cytometer. The results showed that the compound could dose-dependently arrest the cells in the G2 / M phase ( Figure 1 ).

[0272] III. Cell apoptosis experiment

[0273] Test the effect of the compound of the present invention on apoptosis of cancer cells (HT-1080). Seed HT-1080 cells at an appropriate concentration in a 6-well plate and incubate at 37 °C and 5% CO 2 for 24 h, then add different concentrations of the compound or positive control to each well and continue to culture at 37 °C and 5% CO 2 for another 24 h. Collect the cells and centrifuge. Add 500 μL of staining agent (1× Buffer, 5 μL Annexin V-FITC, 5 μL (50 μg / μL) PI) to each sample, stain in the dark at 37 °C for 15 min, centrifuge, remove the dye, wash with PBS, filter through a 300-mesh filter, and detect at 580 nm using a flow cytometer. The results show that the compound can induce apoptosis in a dose-dependent manner ( Figure 2 ).

[0274] IV. Cell colony formation assay

[0275] Test the effect of the compound of the present invention on cell colony formation of cancer cells (HT-1080). Seed HT-1080 cells at an appropriate concentration in a 6-well plate and incubate at 37 °C and 5% CO 2 for 24 h, then add different concentrations of the compound or positive control to each well and continue to culture at 37 °C and 5% CO 2 for 7 days. Discard the culture medium, wash with PBS, fix with 4% paraformaldehyde for 30 min, discard the paraformaldehyde, add 600 μL of 0.1% crystal violet solution to each well, stain for 20 min, wash with water, dry, and take pictures. The results show that the compound can inhibit the proliferation of HT-1080 cells in a dose-dependent manner ( Figure 3 ).

[0276] V. Tubulin polymerization inhibition assay

[0277] Test the inhibitory effect of the compound of the present invention on tubulin polymerization. Use the BK011P kit to test the inhibitory effect of the target compound on tubulin polymerization: prepare the relevant solutions according to the instructions, and then prepare the tubulin polymerization reaction solution required for the experiment by mixing the obtained solutions in an appropriate ratio and place it on ice. Add 5 μL of the test solution to each well of a black 96-well plate and preheat in a microplate reader at 37 °C for 1 minute. After preheating, add 50 μL of the prepared tubulin polymerization reaction solution to each well and record the change in fluorescence intensity per well within 60 minutes. The results show that the compound can inhibit tubulin polymerization in a dose-dependent manner ( Figure 4 ).

[0278] VI. Proliferation inhibition assay of the compound on drug-resistant cells

[0279] The operation procedure is the same as that for the proliferation inhibition assay of tumor cells, and the results are shown in Table 2.

[0280] Table 2. Proliferation inhibition activity of the compound against drug-resistant cells

[0281]

[0282] The experimental results show that the compound inhibits the growth of drug-resistant breast cancer cells, has anti-drug resistance effect, and the anti-drug resistance effect is better than that of colchicine.

[0283] VII. Experiment on the binding of the compound to tubulin

[0284] Test the binding mode of the compound described in the present invention to tubulin. MCF-7 cells were co-incubated with I-19, colchicine or colcemid, and then washed with the culture medium. The cell viability before and after washing was tested. The results show that the binding mode of the compound to tubulin is similar to that of colchicine, different from that of colcemid, and forms a covalent interaction with tubulin, which may be one of the reasons for the anti-drug resistance activity of the compound ( Figure 5 ).

[0285] The above are only the preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. An α, β-unsaturated ketone derivative, characterized in that: The general structural formula of the α,β-unsaturated ketone derivative is shown in formula (I): Among them, R 1 H, -OR 4 、-COOR 4 、-OCOR 4 、-NHCOR 4 or -CONHR 4 , R 1 It can be monosubstituted or disubstituted; R 4 is H, C1-C3 alkyl; R 2 is H, a substituted aromatic ring or a substituted aromatic heterocycle; R 3 is H, Me, Et or CN.

2. The α,β-unsaturated ketone derivative according to claim 1, characterized in that The R 1 is H, -OMe, -OEt, -COOMe, -COOEt, -CONH2, -CONHMe, -NHCOMe or -NHCOEt.

3. The α,β-unsaturated ketone derivative according to claim 2, characterized in that The R 2 for Among them, R 5 is H, F, CH3, t-Bu, CF3, CN, OH, OCH3, NO2, NH2, NHCH3, N(CH3)2 or CONH2, R 5 It may be mono-, di- or tri-substituted.

4. The method for preparing the α,β-unsaturated ketone derivative according to any one of claims 1 to 3, characterized in that: Here are the steps: (1) Compound II is dissolved in anhydrous dichloromethane and reacted with di-tert-butyl dicarbonate in the presence of a base to obtain compound III; wherein the structural formula of compound II is The structural formula of compound III is (2) Compound III is dissolved in methanol, and Pd / C or Raney Ni is used as a catalyst to remove the benzyl group under the action of hydrogen to obtain compound IV; the structural formula of compound IV is (3) Compound IV is dissolved in anhydrous dichloromethane and reacts with acyl chloride in the presence of a base to generate compound V; wherein the structural formula of compound V is (4) Compound V was dissolved in anhydrous dichloromethane, and the Boc protecting group was removed with trifluoroacetic acid to obtain compound I.

5. The method for preparing an α,β-unsaturated ketone derivative according to claim 4, characterized in that: The base is triethylamine or DIPEA; the acyl chloride is Among them, R 3 is H, Me, Et or CN; R 5 is H, F, CH3, t-Bu, CF3, CN, OH, OCH3, NO2, NH2, NHCH3, N(CH3)2 or CONH2, R 5 It may be mono-, di- or tri-substituted.

6. A pharmaceutically acceptable salt of the α,β-unsaturated ketone derivative according to claim 1, characterized in that: The pharmaceutically acceptable salt is an acid addition salt of the compound of general formula (I), wherein the acid used for salt formation is hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or ferulic acid.

7. A pharmaceutical composition, characterized in that The invention comprises the α, β-unsaturated ketone derivative according to claim 1 or the α, β-unsaturated ketone derivative according to claim 6 in a pharmaceutically acceptable salt.

8. Use of the α,β-unsaturated ketone derivative according to claim 1, the pharmaceutically acceptable salt of the α,β-unsaturated ketone derivative according to claim 6, or the pharmaceutical composition according to claim 7 in the preparation of a microtubule polymerization inhibitor.

9. The use according to claim 8, characterized in that: The microtubule polymerization inhibitor is a drug used to treat malignant tumors.

10. The use according to claim 9, characterized in that: The malignant tumor is breast cancer, cervical cancer, fibrosarcoma, ovarian cancer, liver cancer, lung cancer, colon cancer, prostate cancer or chronic myeloid leukemia.

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