A beta-carboline chalcone compound, a preparation method and application thereof

Abstract

The application relates to the field of medicine, and relates to a beta-carboline chalcone compound as well as a preparation method and application thereof. A series of derivatives are synthesized by taking L-tryptophan methyl ester hydrochloride as raw material, a lead compound with good activity is obtained, a new idea is provided for developing a novel YAP1 inhibitor, and it is proved that the derivatives have a strong inhibitory effect on YAP1.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to a β-carboline chalcone compound, its preparation method, and its application. Background Technology

[0002] YAP1 (Yes-associated protein 1) is one of the major factors mediating oncogenes as a driver of gene expression. It has been confirmed to play important roles in organ volume control, stem cell function, tissue regeneration, tumorigenesis, and tumor metastasis. Besides its important role in the homeostasis and regeneration of normal tissues, YAP1 also has significant functions in tumor initiation, invasion, metastasis, and treatment resistance. YAP1 is one of the main effector factors of the Hippo signaling pathway, and it is highly expressed in various solid tumors, regulating tumor cell proliferation, apoptosis, metastasis, and chemoresistance. The transcriptional co-regulator YAP1 is one of the main actors in the Hippo signaling pathway, shuttling between the cytoplasm and the nucleus. Aberrant expression or activation of YAP1 is frequently detected and plays an important role in the progression of various solid tumors, including hepatocellular carcinoma. Emerging evidence suggests that YAP1 is essential in mediating the initiation and growth of most solid tumors. YAP1 primarily regulates malignant tumors by influencing the transcriptional activity of its target genes. YAP1 possesses carcinogenic properties such as promoting proliferation, maintaining stem structure, and inducing treatment resistance. Multiple studies have shown that YAP1 is significantly and abnormally highly expressed in various malignant tumor tissues compared to normal adjacent tissues, and it is also significantly positively correlated with tumor malignancy or invasiveness, closely linked to patient prognosis. Taking ovarian cancer as an example, ovarian cancer is the world's deadliest gynecological malignancy. Although its incidence is lower than other cancers such as endometrial cancer, its mortality rate remains very high. Studies have shown that the pathogenic factors of ovarian cancer include age, infertility, menopause, genetic factors, and unhealthy lifestyle habits. Although the mortality rate of many cancers, such as breast cancer, has been gradually decreasing with the advancement of modern technology, the mortality rate of ovarian cancer has remained unchanged. Currently, ovarian cancer has become a significant public health problem that cannot be ignored; therefore, there is an urgent need to improve treatment methods and diagnostic techniques for ovarian cancer. Summary of the Invention

[0003] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention proposes a β-carbamoline chalcone compound, its preparation method, and its applications.

[0004] In a first aspect, the present invention provides a β-carboline chalcone compound, said β-carboline chalcone compound having a structure as shown in formula (I),

[0005]

[0006] Wherein, R is selected from substituted aromatic groups or heteroaryl groups.

[0007] In some embodiments of the present invention, R includes:

[0008]

[0009] In a second aspect, the present invention provides a method for preparing β-carboline chalcone compounds, comprising: using L-tryptophan methyl ester hydrochloride as a raw material, and synthesizing β-carboline chalcone compounds through cyclization, oxidation, hydrolysis, amide condensation, and aldehyde-ketone condensation reactions.

[0010] In some embodiments of the present invention, the cyclizing agent in the cyclization step includes acetone aldehyde.

[0011] In some embodiments of the present invention, the molar ratio of the cyclizing agent to the L-tryptophan methyl ester hydrochloride is 1.2 to 2:1.

[0012] In some embodiments of the present invention, the cyclization step further includes an acidic catalyst.

[0013] In some embodiments of the present invention, the acidic catalyst comprises sulfuric acid.

[0014] In some embodiments of the present invention, the cyclization step includes: mixing L-tryptophan methyl ester and water, adding acetone aldehyde and sulfuric acid, reacting at room temperature, monitoring the reaction with TLC plate spotting (PE:EA = 2:1), adjusting the pH to neutral, allowing it to stand, and then filtering out the solid to obtain crude product g.

[0015] In some embodiments of the present invention, the cyclization step includes: weighing L-tryptophan methyl ester (7-9 mmol), adding 400 mL of water, stirring at room temperature, adding 577 μL of acetone aldehyde (9-10 mmol), then slowly adding 4 mL of concentrated sulfuric acid, reacting at room temperature for 24-28 h, monitoring the reaction with TLC plate spotting (PE:EA = 2:1), adjusting the pH to 7-8 with NaOH solution, letting it stand for 30-50 min, and filtering out the solid to obtain crude product g.

[0016] In some embodiments of the present invention, the raw materials for the oxidation step include an alkaline catalyst.

[0017] In some embodiments of the present invention, the alkaline catalyst comprises LiOH.

[0018] In some embodiments of the present invention, the oxidation step includes adding LiOH to compound g and a mixed solution (THF:MeOH:H2O = 3:1:1) in an ice bath, heating the mixture for 2-4 hours, and monitoring the reaction using a TLC plate (PE:EA = 2:1). After the reaction is complete, water is added to quench the reaction, and the solid is filtered out to obtain product h.

[0019] In some embodiments of the present invention, the oxidation step includes weighing 500-600 mg of the compound g into a 35 mL sealed tube, adding 4.6 mL of a mixed solution (THF:MeOH:H2O = 3:1:1), adding 156-187.2 mg of LiOH (2-2.4 mmol) under ice bath conditions, mixing well, heating to 50-60 °C, and monitoring the reaction by TLC plate spotting (PE:EA = 2:1) for 2-4 hours. After the reaction is complete, adding approximately 10-20 mL of water to quench the reaction, and then filtering out the solid to obtain product h.

[0020] In some embodiments of the present invention, the raw materials used in the amide condensation step include a condensation reagent.

[0021] In some embodiments of the present invention, the condensing agent comprises 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate.

[0022] In some embodiments of the present invention, the amide condensation step includes mixing product h, HATU, and anhydrous DMF in an ice bath, adding DIPEA and piperidinylpiperidine, purging with nitrogen, reacting at room temperature, monitoring the reaction with TLC plates, quenching with water after the reaction is complete, and filtering the solid to obtain product i.

[0023] In some embodiments of the present invention, the amide condensation step includes: weighing 870–970 mg of product h (3.42 mmol) into a 100 mL two-necked flask, then weighing 1.3–1.5 g of HATU (3.42–3.8 mmol) into the flask, adding 12–20 mL of anhydrous DMF, stirring and mixing under ice bath conditions, then adding 1.2–1.32 mL of DIPEA (6.84–7.6 mmol), and after 25 min, adding 633–700 mg of piperidinylpiperidine (3.76–4.51 mmol), purging with nitrogen three times, reacting at room temperature for 24–28 h, monitoring the reaction by TLC (PE:EA = 2:1), quenching with approximately 10–20 mL of water after the reaction is complete, and filtering the solid to obtain product i.

[0024] In some embodiments of the present invention, the aldehyde-ketone condensation step includes: weighing 120-144 mg of product i (0.3-0.432 mmol) into a 25 mL flask, adding 1 mL of ethanol to dissolve it in an ice bath, then weighing 22-25 mg of KOH (0.39-0.43 mmol) in a small beaker, adding 200-300 μL of water, dissolving it, and then adding it dropwise into a round-bottom flask. Finally, different aldehydes (0.33-0.39 mmol) are added. After 2-3 hours, the reaction is monitored by TLC (DCM:MeOH = 20:1). After the reaction is complete, water is added, the solid is collected by filtration, and then recrystallized with DMF to obtain the final product 1j-35j.

[0025] In a third aspect, the present invention provides an inhibitor for YAP1, said inhibitor having a compound having the structure shown in formula (I).

[0026] A fourth aspect of the invention provides the use of inhibitors in the preparation of therapeutic antitumor drugs.

[0027] In some embodiments of the present invention, the antitumor drug includes an anti-ovarian cancer drug.

[0028] In some embodiments of the present invention, the inhibitor is used to: a) inhibit the proliferation, migration or invasion of ovarian cancer cells; b) promote apoptosis of ovarian cancer cells; or c) arrest the cell cycle of ovarian cancer cells. Attached Figure Description

[0029] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein:

[0030] Figure 1 This invention provides an illustration of the inhibitory effect of YAP1 inhibitor on the stable strain SKOV3-pLV.6YAP1 in Example 3 of the present invention;

[0031] Figure 2 The following are Western blot images of the β-carbazoline chalcone derivatives as YAP1 inhibitors in cells in Example 4 of the present invention: (a) Western blot image of inhibitors (1, 3, 7, 12, 15, 16, 17, 18, 23, 24)j; (b) Western blot image of inhibitors (25, 26, 28, 33, 34)j.

[0032] Figure 3 This is a graph showing the cck8 data of β-carbazoline chalcone derivatives as YAP1 inhibitors against SKOV3 in vivo in Example 5 of the present invention.

[0033] Figure 4 This is the preparation route for β-carbaline chalcone derivatives in the embodiments of the present invention. Detailed Implementation

[0034] The following will describe the concept and technical effects of the present invention clearly and completely with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0035] Example

[0036] Example 1

[0037] This embodiment provides a β-carboline chalcone compound, and the specific preparation method is as follows:

[0038] S1: Synthesis of compound g: Weigh 2g of L-tryptophan methyl ester (8mmol), add 400mL of water, stir at room temperature, add 577uL of acetone aldehyde (9.6mmol), then slowly add 4mL of concentrated sulfuric acid, react at room temperature for 24h, monitor the reaction by TLC (PE:EA=2:1), after the reaction is complete, adjust the pH to 7 with NaOH solution, let stand for 30min, filter out the solid to obtain crude product g.

[0039] S2: Synthesis of compound h: Weigh 500 mg of compound g into a 35 mL sealed tube, add 4.6 mL of mixed solution (THF:MeOH:H2O = 3:1:1), add 156 mg of LiOH (2 mmol) under ice bath, mix well, and heat to 50 °C. After 2 h, monitor the reaction by TLC (PE:EA = 2:1). After the reaction is complete, add about 10 mL of water to quench the reaction and filter out the solid to obtain product h, with a yield of 85%.

[0040] S3: Synthesis of compound i: Weigh 870 mg of product h (3.42 mmol) into a 100 mL two-necked flask, then weigh 1.3 g of HATU (3.42 mmol) into the flask, add 12 mL of anhydrous DMF, stir and mix well under ice bath, then add 1.2 mL of DIPEA (6.84 mmol), and after 25 min, add 633 mg of piperidinylpiperidine (3.76 mmol). Replace with nitrogen three times, react at room temperature for 24 h, and monitor the reaction by TLC (PE:EA = 2:1). After the reaction is complete, add about 10 mL of water to quench the reaction, filter the solid, and obtain product i with a yield of 89%.

[0041] S4: Weigh 120 mg of product i (0.3 mmol) into a 25 mL flask, add 1 mL of ethanol and dissolve in an ice bath. Then weigh 22 mg of KOH (0.39 mmol) into a small beaker, add 200 μL of water, dissolve and add dropwise into a round-bottom flask. Finally, add benzaldehyde (0.33 mmol). After 2 h, monitor the reaction by spotting with a TLC plate (DCM:MeOH = 20:1). After the reaction is complete, add water and filter to collect the solid. Then recrystallize with DMF to obtain the final product 1j.

[0042] Characterization was performed using NMR, MS, and melting point. The structure of the compound is shown below, along with its properties, yield, NMR and mass spectrometry results:

[0043] Compound 1j, molecular formula: C 31 H 32 N4O2, properties: yellow solid, total yield: 41%, melting point: 200-201℃; 1 HNMR (500MHz, CDCl3) δ10.63(s,1H),8.70(s,1H),8.50(d,J=16.1Hz,1H),8.18(d,J=7.8Hz,1H),8.01(d ,J=16.0Hz,1H),7.78-7.73(m,2H),7.63(d,J=3.9Hz,2H),7.49-7.43(m,3H),7.41-7.33(m,1H),4.99-4. 93(m,1H),4.69-4.63(m,1H),3.16-3.07(m,1H),2.92-2.84(m,1H),2.70-2.61(m,1H),2.60(t,J=5.3Hz, 4H),2.04(d,J=13.0Hz,1H),1.96-1.87(m,2H),1.83-1.73(m,1H),1.67-1.59(m,4H),1.51-1.46(m,2H); 13 C NMR (126MHz, CDCl3) δ191.21,167.52,144.38,142.63,141.51,136.53,135.27,133.64,132.43,130.82,129.71,129.11,128.91 ,122.28,121.39,121.08,120.92,120.82,112.33,63.09,50.36,47.59,43.04,29.05,28.07,26.60,24.92; HRMS(ESI-MS)m / z:[C 31 H 32 N4O2+H] +Calculation result for 493.2598; found result for 493.2597. Preparation route as follows: Figure 4 As shown.

[0044] Example 2

[0045] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is p-methylbenzaldehyde, and the reddest product obtained is 2j.

[0046] Compound 2j, molecular formula: C 32 H 34 N4O2, properties: yellow solid, total yield: 37%, melting point: 234-235℃; 1 HNMR (500MHz, CDCl3) δ10.63(s,1H),8.71(s,1H),8.44(d,J=16.0Hz,1H),8.18(d,J=7.9Hz,1H ),7.99(d,J=16.0Hz,1H),7.68-7.60(m,4H),7.41-7.34(m,1H),7.27(s,1H),4.98(d,J=13.2H z,1H),4.71(d,J=13.4Hz,1H),3.15(d,J=12.8Hz,1H),2.94-2.84(m,1H),2.80(s,1H),2.69(s ,4H),2.41(s,3H),2.15-2.00(m,1H),1.99-1.89(m,1H),1.77(d,J=43.9Hz,5H),1.52(s,2H); 13 C NMR (126MHz, CDCl3) δ191.30,167.57,144.48,142.59,141.51,141.40,136.53,133.78,132.61,132.41,129.89,129.70,128.96,12 2.29,121.37,121.12,120.85,119.92,112.32,63.13,50.36,47.60,43.04,29.03,28.08,26.59,24.90,21.76; HRMS(ESI-MS)m / z:[C 32 H 34 N4O2+H] + calcd for 507.2755; found:507.2756.

[0047] Example 3

[0048] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is p-fluorobenzaldehyde, and the reddest product obtained is 3j.

[0049] Compound 3j, molecular formula: C 31 H 31 FN4O2, Properties: Yellow solid, Total yield: 33%, Melting point: 212-213℃; 1 HNMR (500MHz, CDCl3) δ10.61(s,1H),8.69(s,1H),8.40(d,J=16.0Hz,1H),8.18(d,J=7.8Hz,1H),7.96(d,J =16.0Hz,1H),7.77-7.71(m,2H),7.63(d,J=4.0Hz,2H),7.40-7.34(m,1H),7.15(t,J=8.4Hz,2H),4.97(d,J =12.8Hz,1H),4.65(d,J=13.4Hz,1H),3.14(t,J=12.7Hz,1H),2.93-2.84(m,1H),2.75(s,1H),2.66(s,4H), 2.08(s,1H),2.03(d,J=11.3Hz,1H),1.96-1.88(m,1H),1.81-1.75(m,1H),1.70(s,4H),1.57-1.42(m,2H); 13 C NMR (126MHz, CDCl3) δ191.05,167.56,165.26,163.25,143.01,142.65,141.51,136.53,133.62,132.46,131.59,130.85,129.77,122.30 ,121.44,121.07,120.85,120.62,116.39,116.22,112.33,63.05,50.40,47.55,42.97,29.19,27.95,26.52,24.87; HRMS(ESI-MS)m / z:[C 31 H 31 FN4O2+H] + calcd for 511.2504; found:511.2503.

[0050] Example 4

[0051] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is p-chlorobenzaldehyde, and the reddest product obtained is 4j.

[0052] Compound 4j, molecular formula: C 31 H 31 ClN4O2, Properties: Yellow solid, Total yield: 33%, Melting point: 249-250℃; 1 H NMR (500MHz, CDCl3) δ10.59(s,1H),8.69(s,1H),8.45(d,1H),8.18(d,J=7.8Hz,1H),7.94 (d,J=16.0Hz,1H),7.70-7.61(m,4H),7.43(d,J=8.5Hz,2H),7.41-7.34(m,1H),5.00-4.94 (m,1H),4.66-4.60(m,1H),3.13(t,J=12.7Hz,1H),2.93-2.84(m,1H),2.74(s,1H),2.65( s,4H),2.12-2.06(m,1H),1.95-1.86(m,1H),1.81-1.75(m,1H),1.70(s,4H),1.51(s,2H); 13 C NMR (126MHz, CDCl3) δ190.98,167.54,142.75,142.69,141.52,136.67,136.51,133.82,133.55,132.48,130.00,129.79,129.41 ,122.30,121.46,121.43,121.06,120.87,112.34,63.00,50.42,47.57,43.00,29.19,28.06,26.61,24.92; HRMS(ESI-MS)m / z:[C 31 H 31 ClN4O2+H] + calcd for 527.2205; found:527.2208.

[0053] Example 5

[0054] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is p-bromobenzaldehyde, and the reddest product obtained is 5j.

[0055] Compound 5j, molecular formula: C 31 H 31BrN4O2, Properties: Yellow solid, Total yield: 38%, Melting point: 249-250℃; 1H NMR (500MHz, CDCl3) δ 10.60 (s, 1H), 8.69 (s, 1H), 8.46 (d, J = 16.0Hz, 1H), 8.18 (d, J = 7.8Hz, 1H), 7.92 (d, J = 15.9Hz, 1H), 7.68-7.56 (m, 6H), 7.41-7.33 (m, 1H), 4.97 (d, J =13.2Hz,1H),4.64(d,J=13.4Hz,1H),3.13(t,J=12.7Hz,1H),2.93-2.84(m,1H),2. 76(s,3H),2.66(s,4H),2.10(d,J=12.7Hz,1H),2.02(s,1H),1.95-1.84(m,1H); 13C NMR (126MHz, CDCl3) δ191.05,167.58,142.92,142.62,141.54,136.57,134.27,133.59,132.55,132.44,130.23,129.86,125.13 ,122.36,121.59,121.54,121.09,120.94,112.36,63.13,50.39,47.49,42.92,29.03,27.87,26.30,24.74; HRMS(ESI-MS)m / z:[C 31 H 31 BrN4O2+H] + calcd for 572.1737; found:572.1732.

[0056] Example 6

[0057] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is p-cyanobenzaldehyde, and the reddest product obtained is 6j.

[0058] Compound 6j, molecular formula: C 32 H 31 N5O2, properties: yellow solid, total yield: 39%, melting point: 237-238℃; 1HNMR (500MHz, CDCl3) δ10.58(s,1H),8.69(s,1H),8.53(d,J=16.1Hz,1H),8.18(d,J=7.8Hz ,1H),7.95(d,J=16.1Hz,1H),7.83(d,J=8.2Hz,2H),7.76(d,J=8.1Hz,2H),7.69-7.62(m,2 H),7.39(m,1H),5.01-4.94(m,1H),4.62-4.55(m,1H),3.17-3.11(m,1H),2.94-2.85(m,1H ),2.69-2.64(m,4H),2.10(d,J=12.9Hz,1H),1.90-1.84(m,2H),1.72(m,5H),1.52(s,2H); 13 C NMR (126MHz, CDCl3) δ190.62,167.51,142.77,141.55,141.52,139.57,136.56,133.33,132.82,132.61,129.95,129.11,124.08,122 .34,121.62,121.00,120.98,118.53,113.62,112.39,62.94,50.43,47.48,42.88,29.34,27.81,26.45,24.81; HRMS(ESI-MS)m / z:[C 32 H 31 N5O2+H] + calcd for 518.2551; found:518.2548.

[0059] Example 7

[0060] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is p-methoxybenzaldehyde, and the reddest product obtained is 7j.

[0061] Example 8

[0062] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is p-nitrobenzaldehyde, and the reddest product obtained is 8j.

[0063] Example 9

[0064] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is p-sulfonylbenzaldehyde, and the reddest product obtained is 9j.

[0065] Compound 9j, molecular formula: C 32 H 34 N4O4S, properties: yellow solid, total yield: 42%, melting point: 259-260℃; 1 HNMR (500MHz, CDCl3) δ10.57(s,1H),8.68(s,1H),8.61(d,J=16.0Hz,1H),8.19(d,J=7.8Hz,1H),8.03(d,J=8.2Hz,2 H),7.99(d,J=16.1Hz,1H),7.92(d,J=8.2Hz,2H),7.69-7.63(m,2H),7.42-7.35(m,1H),4.95(d,J=13.2Hz,1H),4.5 7-4.50(m,1H),3.15(d,J=12.9Hz,1H),3.10(s,3H),2.93-2.85(m,1H),2.67-2.61(m,1H),2.58(s,4H),2.06(d,J=1 3.0Hz,1H),1.92(d,J=12.1Hz,1H),1.88-1.81(m,1H),1.79-1.71(m,1H),1.63(t,J=5.7Hz,3H),1.51-1.46(m,2H); 13 C NMR (126MHz, CDCl3) δ190.72,167.55,142.91,141.76,141.56,141.40,140.54,136.62,133.37,132.67,129.99,129.40,128.25,12 4.36,122.40,121.67,121.06,121.00,112.39,62.94,50.49,47.53,44.63,42.93,29.38,27.98,26.57,24.89; HRMS(ESI-MS)m / z:[C 32 H 34 N4O4S+H] + calcd for 571.2374; found:571.2369.

[0066] Example 10

[0067] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is p-n-butylbenzaldehyde, and the reddest product obtained is 10j.

[0068] Compound 10j, molecular formula: C 35 H 40N4O2, properties: yellow solid, total yield: 34%, melting point: 233-234℃; 1 HNMR(500MHz, CDCl3)δ10.64(s,1H),8.72(s,1H),8.47(d,J=16.0Hz,1H),8.17(s,1H), 8.01(d,J=16.0Hz,1H),7.70(d,J=8.0Hz,2H),7.63(d,J=4.0Hz,2H),7.49(d,J=8.0Hz,2 H),7.39-7.35(m,,1H),4.98-4.95(m,1H),4.74(s,1H),3.12(t,J=12.7Hz,1H),2.91-2. 86(m,1H),2.70-2.65(m,4H),2.16-1.96(m,3H),1.80(m,1H),1.51(s,2H),1.36(s,9H); 13 C NMR (126MHz, CDCl3) δ191.29,167.58,154.60,144.44,142.47,141.52,136.59,133.77,132.52,132.45,129.75,128.85,126.19,122.3 2,121.42,121.12,120.98,119.89,112.34,63.31,50.33,47.51,42.98,35.16,31.30,28.75,27.91,26.24,24.71; HRMS(ESI-MS)m / z:[C 35 H 40 N4O2+H] + calcd for 549.3224; found:549.3219.

[0069] Example 11

[0070] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is p-trifluoromethylbenzaldehyde, and the reddest product obtained is 11j.

[0071] Compound 11j, molecular formula: C 32 H 31 F3N4O2, Properties: Yellow solid, Total yield: 32%, Melting point: 214-215℃; 1H NMR (500MHz, CDCl3) δ10.58(s,1H),8.70(s,1H),8.57(d,J=16.1Hz,1H),8.18(d,J=7.9Hz,1H),7.99(d ,J=16.1Hz,1H),7.85(d,J=8.1Hz,2H),7.71(d,J=8.0Hz,2H),7.67-7.62(m,2H),7.38(m,1H),4.95(d,J =13.2Hz,1H),4.60(d,J=13.5Hz,1H),3.11(t,J=12.4Hz,1H),2.92-2.84(m,1H),2.58(s,5H),2.05(d,J =13.0Hz,1H),1.97-1.85(m,2H),1.75(d,J=12.4,4.2Hz,1H),1.62(t,J=5.7Hz,4H),1.49-1.45(m,2H); 13 C NMR (126MHz, DMSO) δ189.20,166.39,142.27,141.89,140.79,138.91,134.92,133.15,131.99,130.16,129.90,129.41,129.11,125.80, 125.10,124.08,122.93,122.24,120.78,120.63,120.21,113.33,62.04,49.61,46.71,42.02,27.98,26.04,24.44; HRMS(ESI-MS)m / z:[C 32 H 31 F3N4O2+H] + calcd for 561.2472; found:561.2468.

[0072] Example 12

[0073] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 3-fluorobenzaldehyde, and the reddest product obtained is 12j.

[0074] Example 13

[0075] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 3-chlorobenzaldehyde, and the reddest product obtained is 13j.

[0076] Compound 13j, molecular formula: C 31 H 31ClN4O2, Properties: Yellow solid, Total yield: 33%, Melting point: 233-234℃; 1 H NMR (500MHz, CDCl3) δ10.59(s,1H),8.68(s,1H),8.46(d,J=16.1Hz,1H),8.18(d,J=7.8Hz,1H),7.92( d,J=16.0Hz,1H),7.69(d,J=1.9Hz,1H),7.67-7.61(m,3H),7.44-7.34(m,3H),4.94(d,J=13.1Hz,1H) ,4.60(d,1H),3.18-3.10(m,1H),2.94-2.86(m,1H),2.68-2.60(m,1H),2.57(t,J=5.3Hz,4H),2.04(d ,J=12.9Hz,1H),1.92(d,J=12.3Hz,1H),1.87-1.72(m,1H),1.64-1.58(m,5H),1.46(q,J=5.9Hz,2H); 13 C NMR (126MHz, CDCl3) δ190.91,167.64,142.65,141.54,137.12,136.54,135.10,133.53,132.52,130.57,130.35,129.84,128.91,126 .61,122.31,122.13,121.50,121.04,120.90,112.36,63.09,50.29,47.42,42.88,28.83,27.78,26.23,24.68; HRMS(ESI-MS)m / z:[C 31 H 31 ClN4O2+H] + calcd for527.2208; found:527.2205.

[0077] Example 14

[0078] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 3-bromobenzaldehyde, and the reddest product obtained is 14j.

[0079] Compound 14j, molecular formula: C 31 H 31 BrN4O2, properties: yellow solid, total yield: 32%, melting point: 232-233℃; 1H NMR (500MHz, CDCl3) δ10.58(s,1H),8.68(s,1H),8.46(m,1H),8.18(d,J=7.8Hz,1H),7.90(m,1H) ,7.84(t,J=1.7Hz,1H),7.69(d,J=8.0Hz,1H),7.67-7.60(m,2H),7.59-7.53(m,1H),7.41-7.30(m ,2H),4.94(d,J=13.0Hz,1H),4.56(d,J=13.4Hz,1H),3.20-3.10(m,1H),2.95-2.86(m,1H),2.68 -2.54(m,5H),2.04(m,1H),1.91(m,1H),1.87-1.72(m,1H),1.64-1.57(m,5H),1.50-1.42(m,2H); 13 C NMR (126MHz, CDCl3) δ190.90,167.63,142.79,142.50,141.53,137.41,136.51,133.54,133.46,132.49,132.07,130.54,129.82,126.82 ,123.24,122.31,122.20,121.48,121.04,120.83,112.35,62.95,50.36,47.51,42.96,29.04,28.01,26.59,24.90; HRMS(ESI-MS)m / z:[C 31 H 31 BrN4O2+H]+calcd for 572.1737; found:572.1732.

[0080] Example 15

[0081] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 3-methoxybenzaldehyde, and the reddest product obtained is 15j.

[0082] Compound 15j, molecular formula: C 32 H 34 N4O3, properties: yellow solid, total yield: 43%, melting point: 233-234℃; 1HNMR (500MHz, CDCl3) δ10.61(s,1H),8.70(s,1H),8.44(d,J=16.0Hz,1H),8.18(d,J=7.8Hz,1H ),7.97(d,J=16.0Hz,1H),7.64(d,J=3.8Hz,2H),7.41-7.33(m,3H),7.22(d,J=2.5Hz,1H),7.02 -6.96(m,1H),4.97(d,J=13.2Hz,1H),4.66(d,J=13.4Hz,1H),3.88(s,3H),3.20-3.11(m,1H),2 .94-2.84(m,1H),2.66(s,4H),2.20-1.94(m,2H),1.90-1.75(m,1H),1.71(s,6H),1.49(s,2H); 13 C NMR (126MHz, CDCl3) δ191.23,167.62,160.09,144.39,142.61,141.52,136.66,136.52,133.71,132.44,130.06,129.75,122.29,121.41,12 1.21,121.07,120.85,120.82,116.19,114.88,112.34,63.06,55.52,50.31,47.51,42.96,28.91,27.98,26.39,24.79; HRMS(ESI-MS)m / z:[C 32 H 34 N4O3+H] + calcd for523.2704; found:523.2701.

[0083] Example 16

[0084] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 3-nitrobenzaldehyde, and the reddest product obtained is 16j.

[0085] Compound 16j, molecular formula: C 31 H 31 N5O4, properties: yellow solid, total yield: 39%, melting point: 220-221℃; 1HNMR (500MHz, CDCl3) δ10.57(s,1H),8.68(s,1H),8.61-8.52(m,2H),8.28(d,J=8.2Hz,1H),8.18(d,J =7.9Hz,1H),8.06(d,J=7.7Hz,1H),8.00(d,J=16.0Hz,1H),7.65(t,J=4.5Hz,3H),7.40-7.36(m,1H), 4.94(d,J=13.1Hz,1H),4.53(d,J=13.4Hz,1H),3.19(t,J=12.8Hz,1H),2.92(t,J=13.0Hz,1H),2.67- 2.47(m,4H),2.05(d,J=13.0Hz,1H),1.91(d,J=12.1Hz,1H),1.59(t,J=5.8Hz,4H),1.46-1.43(m,2H); 13 C NMR (126MHz, CDCl3) δ190.57,167.58,148.84,142.83,141.56,141.10,137.06,136.53,133.81,133.35,132.60,130.11,129.94,124.86 ,123.66,123.55,122.32,121.60,121.00,120.93,112.39,62.91,50.37,47.43,42.85,29.15,27.80,26.37,24.75; HRMS(ESI-MS)m / z:[C 31 H 31 N5O4+H]+calcd for538.2449; found:538.2448.

[0086] Example 17

[0087] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 3-cyanobenzaldehyde, and the reddest product obtained is 17j.

[0088] Compound 17j, molecular formula: C 32 H 31 N5O2, properties: yellow solid, total yield: 37%, melting point: 219-220℃; 1HNMR (500MHz, CDCl3) δ10.58(s,1H),8.65(s,1H),8.50(d,J=16.1Hz,1H),8.17(d,J=7.8Hz,1H),7.99(d ,J=7.9Hz,1H),7.96-7.88(m,2H),7.71(d,J=7.7Hz,1H),7.64(d,J=4.0Hz,2H),7.57(t,J=7.8Hz,1H),7 .42-7.34(m,1H),4.97-4.91(m,1H),4.51-4.45(m,1H),3.20-3.12(m,1H),2.95-2.87(m,1H),2.67-2.5 9(m,1H),2.57-2.54(m,4H),2.08-2.02(m,1H),1.91-1.86(m,1H),1.62-1.58(m,5H),1.48-1.43(m,2H); 13 C NMR (126MHz, CDCl3) δ190.66,167.64,142.85,141.56,141.26,136.57,136.54,133.57,133.43,132.64,132.58,131.95,129.95,129.94,123 .23,122.33,121.59,121.00,120.79,118.32,113.58,112.39,62.87,50.38,47.45,42.84,29.19,27.87,26.44,24.81; HRMS(ESI-MS)m / z:[C 32 H 31 N5O2+H] + calcd for 518.2551; found:518.2548.

[0089] Example 18

[0090] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 3-methylbenzaldehyde, and the reddest product obtained is 18j.

[0091] Compound 18j, molecular formula: C 32 H 34 N4O2, properties: yellow solid, total yield: 33%, melting point: 186-187℃; 1HNMR (500MHz, CDCl3) δ10.63(s,1H),8.68(s,1H),8.47(d,J=16.0Hz,1H),8.17(d,J=7.8Hz,1 H),7.99(d,J=16.1Hz,1H),7.67-7.58(m,3H),7.51(s,1H),7.40-7.30(m,2H),4.98-4.91(m,1 H),4.66-4.59(m,1H),3.18-3.09(m,1H),2.94-2.85(m,1H),2.69-2.55(m,5H),2.43(s,3H),2 .07-2.01(m,1H),1.91-1.81(m,1H),1.79-1.72(m,4H),1.65-1.57(m,5H),1.50-1.43(m,3H); 13 C NMR (126MHz, CDCl3) δ191.29,167.68,144.72,142.60,141.52,138.81,136.52,135.22,133.81,132.41,131.68,130.37,129.72,128.97,12 5.39,122.28,121.38,121.08,120.75,120.65,112.33,63.07,50.31,47.51,42.96,28.96,27.90,26.43,24.81,21.54; HRMS(ESI-MS)m / z:[C 32 H 34 N4O2+H]+calcd for507.2755; found:507.2752.

[0092] Example 19

[0093] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 2-fluorobenzaldehyde, and the reddest product obtained is 19j.

[0094] Compound 19j, molecular formula: C 31 H 31 FN4O2, Properties: Yellow solid, Total yield: 38%, Melting point: 225-226℃; 1H NMR (500MHz, CDCl3) δ10.60(s,1H),8.70(s,1H),8.47(d,J=16.0Hz,1H),8.18(d,J=7.8Hz,1H),7 .94(d,J=16.0Hz,1H),7.68-7.60(m,2H),7.51(d,J=7.6Hz,1H),7.46-7.33(m,3H),7.18-7.10(m, 1H),4.98-4.92(m,1H),4.66-4.60(m,1H),3.16-3.08(m,1H),2.93-2.85(m,1H),2.69-2.60(m,1H ),2.60-2.57(m,4H),2.07-2.01(m,1H),1.97-1.73(m,3H),1.65-1.58(m,5H),1.49-1.44(m,2H); 13 C NMR (126MHz, CDCl3) δ191.22,167.50,163.03,161.00,142.70,141.51,136.84,136 .82,136.57,133.51,132.48,132.19,132.12,129.79,129.76,124.65,124.63,123 .47,123.38,123.29,123.24,122.30,121.44,121.08,121.05,116.51,116.33,112 .34,63.06,50.29,47.61,43.05,29.08,27.91,26.62,24.95; HRMS(ESI-MS)m / z:[C 31 H 31 FN4O2+H] + calcd for 511.2504; found:511.2502.

[0095] Example 20

[0096] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 2-chlorobenzaldehyde, and the reddest product obtained is 20j.

[0097] Compound 20j, molecular formula: C 31 H 31 ClN4O2, Properties: Yellow solid, Total yield: 35%, Melting point: 221-222℃; 1H NMR (500MHz, CDCl3) δ10.62(s,1H),8.69(s,1H),8.51-8.40(m,2H),8.18(d,J=7.8Hz,1 H),7.90(d,J=7.1Hz,1H),7.64(d,J=4.3Hz,2H),7.48(d,J=7.4Hz,1H),7.37-7.34(m,3H ),4.97-4.91(m,1H),4.63-4.57(m,1H),2.87(t,J=12.8Hz,1H),2.62(s,2H),2.55(d,J= 6.0Hz,4H),2.04-2.00(m,1H),1.91-1.87(m,1H),1.63-1.59(m,5H),1.49-1.41(m,2H); 13 C NMR (126MHz, CDCl3) δ190.93,167.55,142.67,141.55,140.02,136.59,136.03,133.55,132.52,131.47,130.49,129.83,127.97 ,127.25,123.22,122.32,121.50,120.94,112.37,63.05,50.32,47.58,42.99,29.07,27.93,26.50,24.85; HRMS(ESI-MS)m / z:[C 31 H 31 ClN4O2+H] + calcd for 527.2208; found:527.2206.

[0098] Example 21

[0099] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 2-bromobenzaldehyde, and the reddest product obtained is 21j.

[0100] Compound 21j, molecular formula: C 31 H 31 BrN4O2, properties: yellow solid, total yield: 32%, melting point: 224-225℃; 1H NMR (500MHz, CDCl3) δ10.63(s,1H),8.69(s,1H),8.45-8.34(m,2H),8.17(d,J=7.8Hz,1H),7.88(d, J=7.7Hz,1H),7.69-7.61(m,3H),7.44-7.34(m,2H),7.27(s,1H),4.95(d,J=13.0Hz,1H),4.61(d,J =13.3Hz,1H),3.11(t,J=12.6Hz,1H),2.87(t,J=12.8Hz,1H),2.71-2.63(m,1H),2.59(s,4H),2.05 (d,J=12.6Hz,1H),1.94(d,J=9.6Hz,1H),1.78-1.75(m,1H),1.66-1.62(m,3H),1.50-1.44(m,2H); 13 C NMR (126MHz, CDCl3) δ190.84,167.54,142.64,141.55,136.58,135.29,133.75,133.49,132.51,131.61,129.82,128.10,127.87,126 .53,123.40,122.31,121.48,121.05,120.93,112.37,63.03,50.30,47.58,42.98,29.05,27.93,26.51,24.85; HRMS(ESI-MS)m / z:[C 31 H 31 BrN4O2+H]+calcd for 572.1737; found:572.1732.

[0101] Example 22

[0102] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the reddest product obtained in step S4 is 22j, where the aldehyde is 2-nitrobenzaldehyde.

[0103] Compound 22j, molecular formula: C 32 H 34 N4O3, properties: yellow solid, total yield: 43%, melting point: 222-223℃; 1HNMR (500MHz, CDCl3) δ10.60(s,1H),8.68(s,1H),8.41(s,2H),8.18(d,J=7.9Hz,1H),8.09(d,J= 8.2Hz,1H),7.90(d,J=7.7Hz,1H),7.75(t,J=7.6Hz,1H),7.67-7.63(m,2H),7.59(t,J=8.0Hz,1H ),7.41-7.35(m,1H),4.94(d,J=13.1Hz,1H),4.54(d,J=13.4Hz,1H),3.10(t,J=12.9Hz,1H),2.8 6(t,J=12.9Hz,1H),2.67(s,1H),2.56(s,4H),2.04(d,J=12.6Hz,1H),1.62(s,4H),1.46(s,2H); 13 C NMR (126MHz, CDCl3) δ190.41,167.57,149.05,142.71,141.60,139.29,136.62,133.67,133.32,132.62,131.62,130.53,129. 96,129.43,125.47,125.19,122.34,121.60,121.03,120.93,112.44,62.99,50.28,47.50,42.87,29.07,27.78,26.35,24.75.

[0104] Example 23

[0105] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 2-methoxybenzaldehyde, and the reddest product obtained is 23j.

[0106] Compound 23j, appearance: yellow solid, overall yield: 31%, melting point: 207-208℃; 1H NMR (500MHz, CDCl3) δ10.66 (s, 1H), 8.70 (s, 1H), 8.45 (s, 2H), 8.18 (d, J = 7.8Hz, 1H), 7.84-7. 79(m,1H),7.63(d,J=3.7Hz,2H),7.45-7.32(m,2H),7.03(t,J=7.5Hz,1H),6.97(d,J=8.3Hz, 1H),4.97(d,J=13.2Hz,1H),4.68(d,J=13.5Hz,1H),3.95(s,3H),3.14(t,J=12.7Hz,1H),2.9 3-2.83(m,1H),2.74(s,2H),2.64(s,4H),2.12-2.05(m,2H),1.83-1.72(m,1H),1.49(s,2H); 13 C NMR (126MHz, CDCl3) δ191.50,167.65,158.98,142.52,141.51,139.36,136 .57,133.93,132.38,132.26,129.67,128.11,124.40,122.29,121.34,121. 14,120.91,120.82,120.78,112.33,111.47,77.41,77.16,76.91,63.15,55 .80,50.31,47.56,42.99,28.94,27.96,26.41,24.80; HRMS(ESI-MS)m / z:[C 32 H 34 N4O3+H] + calcd for 523.2704; found:523.2701.

[0107] Example 24

[0108] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 2-methylbenzaldehyde, and the reddest product obtained is 24j.

[0109] Example 25

[0110] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 2-pyridinecarboxaldehyde, and the reddest product obtained is 25j.

[0111] Compound 25j, molecular formula: C 30 H 31 N5O2, properties: yellow solid, total yield: 31%, melting point: 195-196℃;1 HNMR(500MHz, CDCl3)δ10.60(s,1H),8.87(d,J=15.8Hz,1H),8.73(d,J=3.6Hz,2H),8.1 8(d,J=7.8Hz,1H),7.99(d,J=15.8Hz,1H),7.81-7.74(m,1H),7.67-7.61(m,3H),7.42-7 .35(m,1H),7.35-7.29(m,1H),5.00-4.93(m,1H),4.75-4.69(m,1H),3.16(d,J=12.1Hz, 1H),2.93-2.84(m,1H),2.67(s,5H),2.21-1.96(m,3H),1.81-1.75(m,1H),1.50(s,2H); 13 CNMR(126MHz, CDCl3)δ191.57,167.44,153.84,150.46,142.97,142.69,141.52,136.83,136.54,133.52,132.49,129.77,124.86 ,124.78,124.45,122.30,121.47,121.09,112.33,63.16,50.25,47.64,43.03,29.00,27.82,26.44,24.83; HRMS(ESI-MS)m / z:[C 32 H 34 N4O2+H] + calcd for 494.2551; found:494.2545.

[0112] Example 26

[0113] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 3-pyridinecarboxaldehyde, and the reddest product obtained is 26j.

[0114] Compound 26j, molecular formula: C 30 H 31 N5O2, properties: yellow solid, total yield: 29%, melting point: 233-234℃; 1HNMR (500MHz, CDCl3) δ10.61(s,1H),8.96(d,J=2.2Hz,1H),8.73(s,1H),8.71-8.66(m,1H),8.57(d,J= 16.2Hz,1H),8.21(d,J=7.8Hz,1H),8.14-8.08(m,1H),8.01(d,J=16.1Hz,1H),7.67(d,J=4.0Hz,2H),7. 49-7.42(m,1H),7.42-7.37(m,1H),5.01(d,J=13.1Hz,1H),4.66(d,J=13.4Hz,1H),3.20(t,J=12.7Hz,1 H),2.93(t,J=11.2Hz,1H),2.71(s,4H),2.15(d,1H),1.91(d,J=11.7Hz,1H),1.83(s,2H),1.76(s,7H); 13 C NMR (126MHz, CDCl3) δ190.65,167.52,151.30,150.63,142.70,141.56,140.33,136.57,134.72,133.37,132.59,131.08,129.89,123 .97,122.85,122.34,121.56,121.04,120.99,112.38,62.95,50.37,47.52,42.94,29.24,27.80,26.37,24.78; HRMS(ESI-MS)m / z:[C 30 H 31 N5O2+H]+calcd for494.2551; found:494.2544.

[0115] Example 27

[0116] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is p-dimethylaminobenzaldehyde, and the reddest product obtained is 27j.

[0117] Example 28

[0118] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the reddest product obtained in step S4 is 28j, where the aldehyde is 3,4,5-3-methoxybenzaldehyde.

[0119] Compound 28j, molecular formula: C 34 H 38 N4O5, properties: yellow solid, total yield: 36%, melting point: 233-234℃;1 HNMR (500MHz, CDCl3) δ10.61(s,1H),8.64(s,1H),8.33(d,J=15.9Hz,1H),8.18(d,J=7.9Hz,1H) ,7.94(d,J=16.0Hz,1H),7.68-7.61(m,2H),7.42-7.34(m,1H),6.97(s,2H),4.94(d,J=13.1Hz, 1H),4.47(d,J=13.3Hz,1H),3.97(s,6H),3.92(s,3H),3.18(t,J=12.7Hz,1H),2.91(t,J=12.6H z,1H),2.57(s,5H),2.10(s,1H),1.92(s,1H),1.72(d,J=12.5Hz,2H),1.63(s,6H),1.45(s,2H); 13 C NMR (126MHz, CDCl3) δ191.16,167.98,153.64,144.65,142.75,141.53,140.77,136.46,134.09,132.42,130.88,129.79,122.29,121.4 2,121.07,120.38,120.25,112.34,106.20,62.87,61.16,56.52,50.46,47.44,42.75,29.46,27.91,26.35,24.78; HRMS(ESI-MS)m / z:[C 34 H 38 N4O5+H] + calcd for583.2915; found:583.2910.

[0120] Example 29

[0121] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the reddest product obtained in step S4 is 29j, where the aldehyde is 3,4,5-3-fluorobenzaldehyde.

[0122] Compound 29j, molecular formula: C 31 H 29 F3N4O2, Properties: Yellow solid, Total yield: 29%, Melting point: 244-245℃; 1H NMR (500MHz, CDCl3) δ10.55(s,1H),8.68(s,1H),8.38(d,J=16.0Hz,1H),8.18( d,J=7.9Hz,1H),7.80(d,J=16.0Hz,1H),7.66-7.61(m,2H),7.41-7.31(m,3H),5 .00-4.92(m,1H),4.61-4.55(m,1H),3.15(t,J=12.7Hz,1H),2.90(t,J=12.5Hz, 1H),2.67(d,J=43.8Hz,5H),2.14-1.93(m,2H),1.85-1.74(m,3H),1.49(s,2H); 13 C NMR (126MHz, CDCl3) δ190.54,167.57,142.91,141.55,140.77,136.59,133.31,132.63,131.54,129.96,122.93,122.38,121.63 ,121.05,121.02,112.72,112.68,112.55,112.37,63.02,50.40,47.49,42.95,28.98,28.03,26.45,24.80; HRMS(ESI-MS)m / z:[C 31 H 29 F3N4O2+H] + calcd for547.2315; found:547.2311.

[0123] Example 30

[0124] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the aldehyde in step S4 is 2-naphthoaldehyde, and the reddest product obtained is 30j.

[0125] Compound 30j, molecular formula: C 35 H 34 N4O2, properties: yellow solid, total yield: 37%, melting point: 208-209℃; 1HNMR (500MHz, CDCl3) δ10.66(s,1H),8.70(s,1H),8.58(d,J=15.9Hz,1H),8.16(d,J=16.1Hz, 2H),8.10(s,1H),7.93-7.84(m,4H),7.63(d,J=3.9Hz,2H),7.56-7.52(m,2H),7.38-7.34(m,1 H),4.98(d,J=13.0Hz,1H),4.69(d,J=13.3Hz,1H),3.13(d,J=13.2Hz,1H),2.90(t,J=12.6Hz ,1H),2.62(s,4H),2.08(s,1H),1.96(d,J=15.7Hz,1H),1.66-1.62(m,4H),1.49-1.45(m,2H); 13 C NMR (126MHz, CDCl3) δ191.22,167.65,144.51,142.65,141.54,136.56,134. 59,133.79,133.53,132.83,132.46,131.59,129.76,128.93,128.88,127.97 ,127.63,126.94,123.78,122.32,121.43,121.11,121.05,120.86,112.34,6 3.10,50.38,47.62,43.06,29.06,28.06,26.56,24.89; HRMS(ESI-MS)m / z:[C 35 H 34 N4O2+H]+calcdfor 543.2755; found:543.2750.

[0126] Example 31

[0127] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the reddest product obtained in step S4 is 31j, where the aldehyde is 3-trifluoromethylbenzaldehyde.

[0128] Compound 31j, molecular formula: C 32 H 31 F3N4O2, Properties: Yellow solid, Total yield: 28%, Melting point: 204-205℃; 1H NMR (500MHz, CDCl3) δ10.59(s,1H),8.68(s,1H),8.52(d,J=16.1Hz,1H),8.18(d,J=7.9Hz,1H),8.00(d,J= 16.1Hz,1H),7.96(d,J=7.8Hz,1H),7.92(s,1H),7.69(d,J=7.8Hz,1H),7.64(d,J=4.0Hz,2H),7.63-7.56( m,1H),7.42-7.33(m,1H),4.98-4.92(m,1H),4.61-4.55(m,1H),3.18(t,J=12.6Hz,1H),2.96-2.87(m,1H) ,2.69(s,1H),2.60(s,4H),2.07(d,J=12.8Hz,1H),1.95(s,1H),1.86-1.74(m,2H),1.47(d,J=8.2Hz,2H); 13 CNMR(126MHz, CDCl3)δ190.91,167.68,142.80,142.39,141.56,136.56,136.12,133.55,132.58,131.83,131.57,131.04,129.90,129.63,12 7.09,126.13,122.63,122.35,121.56,121.06,120.86,112.37,62.90, 50.34,47.49,42.93,29.08,27.91,26.49,24.84; HRMS(ESI-MS)m / z:[C 32 H 31 F3N4O2+H]+calcd for 561.2472; found:561.2470.

[0129] Example 32

[0130] This embodiment provides a β-carbamoline chalcone compound. The difference between this embodiment and Example 1 is that the reddest product obtained in step S4 is 32j, where the aldehyde is 2-trifluoromethylbenzaldehyde.

[0131] Compound 32j, molecular formula: C 32 H 31 F3N4O2, Properties: Yellow solid, Total yield: 33%, Melting point: 233-234℃; 1H NMR (500MHz, CDCl3) δ10.61(s,1H),8.69(s,1H),8.45(d,J=15.8Hz,1H),8.38(d,J=15.9,2.5Hz,1H),8.1 8(d,J=7.9Hz,1H),7.99(d,J=7.8Hz,1H),7.76(d,J=7.8Hz,1H),7.70-7.60(m,3H),7.57-7.50(m,1H),7. 41-7.34(m,1H),4.98-4.92(m,1H),4.62-4.56(m,1H),3.15-3.07(m,1H),2.91-2.82(m,1H),2.69(s,1H) ,2.58(s,4H),2.05(d,J=12.7Hz,1H),1.94(s,1H),1.88-1.68(m,2H),1.64(s,5H),1.47(d,J=8.4Hz,2H); 13 C NMR (126MHz, CDCl3) δ190.65,167.55,142.67,141.57,139.53,136.62,134.28,133.41,132.57,132.27,129.92,129.73,129.49,128.20,126 .47,125.25,124.71,123.08,122.33,121.55,121.00,112.38,63.05,50.28,47.52,42.91,29.01,27.79,26.37,24.76.HRMS(ESI-MS)m / z:[C 32 H 31 F3N4O2+H]+calcd for 561.2472; found:561.2465.

[0132] Test Example 1

[0133] Construct a stable GFP-YAP1 strain, SKOV3-pLV.6-YAP1.

[0134] 1. Preparation of reagents and standard solutions:

[0135] (1) Preparation of DMEM cell culture medium: Take two packets of DMEM powder and add them to 1600mL of sterile water. Then weigh 7.4g sodium bicarbonate, 0.2g penicillin and 0.2g streptomycin. Stir well and bring the volume to 2000mL. Filter the solution through a 0.22μm filter membrane in a biosafety cabinet and collect it in a sterile bottle. Seal and store at 4℃ for later use.

[0136] (2) Preparation of 10xPBS solution:

[0137] Weigh 80g of sodium chloride, 2g of potassium chloride, 2.4g of potassium dihydrogen phosphate, and 35.7g of disodium hydrogen phosphate dodecahydrate into a 2L beaker, add 800mL of double-distilled water, and after complete dissolution, adjust the pH to about 7.4, bring the volume to 1L, dispense into glass bottles for later use, and dilute to 1x with sterile ultrapure water when needed.

[0138] (3) Cell culture: Ovarian cancer cells SKOV3 were cultured in DMEM medium containing 10% FBS under the following conditions: 37°C, 5% carbon dioxide. Cells were passaged every other day, and healthy ovarian cancer cells were used for subsequent experiments.

[0139] 2. Experimental Procedures: (Steps for constructing stable YAP1 strains):

[0140] 1. Virus Packaging

[0141] 1. Prepare 6cm 293T cells with a density of 80%-90% in advance.

[0142] 2. Replace the 293T cells in DMEM medium containing 10% FBS with DMEM medium without FBS.

[0143] 3. Take a sterile EP tube, label it with the name, add 600μL opti-MEM and 18μL PEI, mix well and let stand for 5min.

[0144] 4. Add 2.25 μg PSPAX3, 0.75 μg PMD2G and 3 μg target plasmid, mix and let stand for 20 min.

[0145] 5. The above mixture was added dropwise into 293T cells.

[0146] 6. After 8-12 hours, replace with 5 mL of culture medium containing serum.

[0147] 7. Collect the first viral fluid after 48 hours. Collect the 293T cell culture medium, filter it through a 0.45μm filter membrane, store it at -80℃ for later use, and then add 5mL of culture medium containing serum.

[0148] 8. Collect the second batch of virus solution after 72 hours and store it at -80℃ for later use.

[0149] 2. Viral infection and stable strain construction:

[0150] 1. Take cells in good growth condition, discard the culture medium, add PBS to wash away the residual culture medium, add 500 μL of trypsin for digestion, add 1 mL of culture medium to stop digestion, centrifuge at 800 rpm for 5 min, discard the supernatant, add 1 mL of culture medium to resuspend, take 10 μL of the suspension, add 90 μL of PBS and mix well, then take 10 μL of the mixture for counting.

[0151] 2. Lay 4 x 10 5 One cell was placed in a 6cm cell culture dish, and the culture medium in the dish was added to make up to 3mL.

[0152] 3. After 24 hours of plating, discard the culture medium, add 1.5 mL of fresh culture medium, then add 1.5 mL of virus solution, then add 30 μL of Hepes (1 M Hepes: culture medium = 1:100), then add 3 μL of polybrene (polybrene: culture medium = 1:1000), mix, and then place in an incubator for incubation.

[0153] 4. Change the medium after 12 hours, add normal culture medium, and continue culturing for 8-10 hours before secondary infection.

[0154] 5. After the secondary infection, the cells were passaged after reaching confluence, and 3 μL of puromycin was added (puromycin: medium = 1:1000).

[0155] 6. Three days after screening with puromycin, a portion of the cells can be collected for Western blot verification to confirm whether the stable cell line has been successfully constructed.

[0156] Example 3: Screening of small molecule compounds:

[0157] 1. Preparation of reagents and standard solutions:

[0158] (1) Preparation of DMEM cell culture medium: Take two packets of DMEM powder and add them to 1600mL of sterile water. Then weigh 7.4g sodium bicarbonate, 0.2g penicillin and 0.2g streptomycin. Stir well and bring the volume to 2000mL. Filter the solution through a 0.22μm filter membrane in a biosafety cabinet and collect it in a sterile bottle. Seal and store at 4℃ for later use.

[0159] (2) Preparation of 10x PBS solution:

[0160] Weigh 80g of sodium chloride, 2g of potassium chloride, 2.4g of potassium dihydrogen phosphate, and 35.7g of disodium hydrogen phosphate dodecahydrate into a 2L beaker, add 800mL of double-distilled water, and after they are completely dissolved, adjust the pH to about 7.4, bring the volume to 1L, and dispense into glass bottles for later use.

[0161] (3) Cell culture: Stable line SKOV3-pLV.6-YAP1 was cultured in DMEM medium containing 10% FBS under the following conditions: 37℃, 5% CO2. Cells were passaged every other day, and healthy ovarian cancer cells were used for subsequent experiments.

[0162] (4) Compound dilution solution: Dilute the compound to a concentration of 10 mM using DMSO. Take 1.1 μL and add it to 100 μL of medium containing 10% FBSDMEM.

[0163] 2 Experimental Procedure

[0164] 1. Place 16,000 stable strains of SKOV3-pLV.6-YAP / well, 100 μL, into a 96-well blackboard.

[0165] 2. After the cells have adhered and stabilized, add 10 μL of the compound dilution buffer for treatment. Set up three replicates for each inhibitor.

[0166] 3. After 24 hours, remove the culture medium, add 100 μL of PBS, and use a high-throughput multimode microplate detection system to measure the fluorescence value.

[0167] 1. The experimental results are shown in Table 2.

[0168] Table 2. Evaluation of the inhibitory activity of the screened compound YAP1

[0169]

[0170]

[0171]

[0172] Results Analysis: From Table 2 and Figure 1 It is clear that the compound at a final concentration of 10 μM can reduce the fluorescence intensity of stable cell lines. Among them, the compounds with significant effects are (1j, 3j, 7j, 12j, 15j, 16j, 17j, 18j, 23j, 24j, 25j, 26j, 28j, 33j, 34j), with values ​​around 0.5.

[0173] Test Example 2: Western Blotting of Proteins

[0174] 1. Preparation of reagents and standard solutions

[0175] (1) Preparation of 6x Loading buffer: Weigh 4.8g of 10% SDS, then add 2.88mL of 1M Tris-HCl (pH 6.8), 8.8mL of 1% bromophenol blue, 2.4mL of β-mercaptoethanol, 20mL of 50% glycerol, and 5.92mL of double-distilled water into a 50mL centrifuge tube. Mix thoroughly and place in a shaker at 37℃ overnight. Store at -20℃ the next day for later use.

[0176] (2) Preparation of 20x TBST: Weigh 121.4g of Tris and 175.0g of sodium chloride into a 2L beaker, add 40mL of Tween20 and 800mL of double-distilled water. After stirring and dissolving, adjust the pH to 8.0 with concentrated hydrochloric acid or 5M sodium hydroxide, and then bring the volume to 1L with double-distilled water. Store at room temperature for later use.

[0177] (3) Preparation of 1x transfer solution: Weigh 3.03g of Tris and 14.41g of glycine and dissolve them in 800mL of double-distilled water. After the solution is completely dissolved by stirring, add 200mL of methanol, mix well, store at room temperature for later use, and reuse no more than three times.

[0178] (4) Preparation of 1x electrophoresis: Weigh 3.03g of Tris, 18.76g of glycine and 1g of SDS powder and dissolve them in 1000mL of double-distilled water. After complete stirring and dissolution, store at room temperature for later use. It can be reused no more than three times.

[0179] (5) Preparation of 5% skim milk: Weigh 10g of skim milk powder and add 200mL of 1x TBST. After it is completely dissolved by stirring, add 200μL of sodium azide, mix well, and store at 4℃ for later use.

[0180] (6) Spread SKOV3 cells onto a plate. When the cell density is 80-90%, replace the plate with 3 mL of 10% FBS and add 3 μL of 10 mM compound (1j, 3j, 7j, 12j, 15j, 16j, 17j, 18j, 23j, 24j, 25j, 26j, 28j, 33j, 34j) for 24 h.

[0181] 2 Experimental Procedure

[0182] 1. Collect cells, add pre-chilled PBS to wash away residual culture medium, add 1 mL pre-chilled PBS, scrape the cells off on ice with a cell scraper, collect them into 1.5 mL EP tubes, centrifuge at 3000 rpm for 5 min, discard the supernatant, add lysis buffer according to the cell volume, and sonicate (10% intensity, sonicate for 5 seconds, stop for 3 seconds, sonicate for a total of 30 seconds).

[0183] 2. Centrifuge at 12000 rpm for 10 min at 4℃. Discard the precipitate and keep the supernatant. Transfer the supernatant to a clean EP tube and add 6x loading buffer.

[0184] 3. Heat the sample in a 95℃ metal bath for 5 minutes.

[0185] 4. Loading the sample: Place the gel plate into the electrophoresis tank. Put 2 μL of marker in the first well, then put the protein sample in the next well, and put 1x loading buffer of the same volume as the protein sample in the last well.

[0186] 5. Electrophoresis, 100V 90min. Wet transfer: After electrophoresis, remove the gel, discard the compression gel, and retain the separating gel. Pour 1x transfer buffer into the tray and place the gel in the following order: blackboard clip-sponge-filter paper-gel-PVDF membrane-filter paper-sponge-white board. Gently remove air bubbles with a roller. Then, place the blackboard clip into the transfer tank with the negative electrode facing the tank. Perform the transfer at 100V 90min. The PVDF membrane needs to be activated with 100% methanol before use.

[0187] 6. After the transfer is complete, remove the membrane and wash it three times with 1x TBST for 10 minutes each time.

[0188] 7. Seal the membrane and place it in 5% skim milk. Incubate at room temperature for 1 hour.

[0189] 8. Incubate the primary antibody. Prepare the antibody according to the instructions and incubate overnight on a shaker at 4°C.

[0190] 9. Recover the primary antibody, wash three times with 1x TBST, 10 min each time.

[0191] 10. Incubate the secondary antibody. The secondary antibody is prepared according to the properties of the primary antibody and incubated at room temperature for 1 hour.

[0192] 11. Recover the secondary antibody, wash three times with 1x TBST, 10 min each time.

[0193] 12. Development: Use an ultra-sensitive multi-functional imager for development. Dry the PVDF film with TBST, add the luminescent liquid, and place it in the instrument to develop the bands.

[0194] 3 Results Analysis

[0195] Based on the fluorescence values ​​detected by high-throughput multimode microplate testing, compounds that significantly reduced fluorescence were selected for Western blotting experiments. The selected compounds were (1j, 3j, 7j, 12j, 15j, 16j, 17j, 18j, 23j, 24j, 25j, 26j, 28j, 33j, 34j). Figure 2It can be seen that the YAP1 protein bands that are significantly reduced are (1j, 3j, 16j, 17j, 18j, 25j, 26j, 28j, 33j, 34j), indicating that these compounds can significantly reduce the YAP1 protein level in SKOV3.

[0196] Test Example 3: CCK8 Experiment

[0197] 1. Preparation of reagents and standard solutions

[0198] (1) Preparation of DMEM cell culture medium: Take two packets of DMEM powder and add them to 1600mL of sterile water. Then weigh 7.4g sodium bicarbonate, 0.2g penicillin and 0.2g streptomycin. Stir well and bring the volume to 2000mL. Filter the solution through a 0.22μm filter membrane in a biosafety cabinet and collect it in a sterile bottle. Seal and store at 4℃ for later use.

[0199] 2 Experimental Procedure

[0200] The following steps were performed using the purchased CCK8 reagent kit: (1) Plating: 2000 SKOV3 cells were plated in each well of a 96-well plate, and 100 μL of culture medium was added to each well. (2) Adding the drug after 24 hours: The concentration gradient was set according to the experimental requirements, and 50 μL was added to each well. (3) After 72 hours, 15 μL of CCK8 was added to each well using a multipipeline. (4) After incubation at 37°C for 1 hour, the OD value at 450 nm was detected using a microplate reader. (5) Data processing: Cell viability was calculated (cell viability = [(As-Ab) / (Ac-Ab)] x 100%, where As is the absorbance of the experimental well, Ac is the absorbance of the control well, and Ab is the absorbance of the control well).

[0201] 3 Results Analysis

[0202] Compounds (1j, 3j, 16j, 17j, 18j, 25j, 26j, 28j, 33j, 34j) that can significantly reduce the YAP1 protein level of SKOV3 were selected for CCK8 assay. Figure 3 It can be clearly seen that the IC50 values ​​of the compounds are in the range of 0.1-2 μM, with compound 25j exhibiting the best activity and an IC50 of 0.1352 μM. This series of compounds shows strong toxicity to ovarian cancer cells and can significantly reduce YAP1 protein levels.

[0203] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. A β-carboline chalconoid compound, characterized in that, The β-carboline chalcone compounds are compounds having the structure shown in formula (I). Formula (I); Wherein, R is selected from one of the following structures: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 2. The method for preparing β-carboline chalcone compounds according to claim 1, characterized in that: The preparation route of the β-carboline chalcone compounds is shown below: ； In step i, the cyclizing agent is acetone aldehyde; In step ii, the raw materials include an alkaline catalyst, wherein the alkaline catalyst is LiOH; In step iii, the raw materials include a condensation reagent, which is 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate.

3. An inhibitor against YAP1, characterized in that, The inhibitor has the β-carboline chalcone compound as described in claim 1.

4. The use of the inhibitor as described in claim 3 in the preparation of an antitumor drug; wherein the antitumor drug is an anti-ovarian cancer drug.

5. Use according to claim 4, characterized in that, The inhibitors are used to: a) inhibit the proliferation, migration, or invasion of ovarian cancer cells; b) promote apoptosis of ovarian cancer cells; or c) arrest the cell cycle of ovarian cancer cells.

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