Pyridine dimethoxy phenyl semicarbazide derivative as well as preparation method and application thereof
By developing pyridine bimethoxyphenylinide derivatives, the problems of low activity and high synthesis cost of small molecule compounds have been solved, effective inhibition and production efficiency of pancreatic cancer cells have been achieved, and more effective for pancreatic cancer patients have been provided. treatment options.
Patent Information
- Application Number
- CN202411403281.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-09
- Publication Date
- 2025-01-10
AI Technical Summary
Existing small molecule compounds have low activity in the treatment of pancreatic cancer, cumbersome and high cost, which limits their accessibility and market application in clinical applications.
A pyridine bimethoxyphenylinide derivative was developed, which was synthesized through condensation reaction, with good activity that can induce apoptosis, inhibit the growth of pancreatic cancer cells, and optimize the synthesis route to reduce production costs.
Effective inhibition of different pancreatic cancer cells is achieved, production costs are reduced, synthesis efficiency is improved, and more effective and economical treatment options are provided.
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Abstract
Description
Technical Field
[0001] The invention relates to the technical field of drug preparation, and in particular to a pyridine dimethoxyphenylaminourea derivative and a preparation method and application thereof. Background Art
[0002] Pancreatic cancer is a malignant tumor that is mainly divided into adenocarcinoma, endocrine tumors and other rare types, among which adenocarcinoma accounts for the majority of cases. According to statistics, the incidence of pancreatic cancer has been increasing year by year worldwide, especially in North America and Europe, and the incidence in China is also increasing. According to the latest research, the global mortality rate of pancreatic cancer ranks first among all cancers, causing hundreds of thousands of deaths each year. Although scientists have improved their understanding of the disease, the five-year survival rate is still very low, only about 10%. This is mainly attributed to the lack of obvious early symptoms of pancreatic cancer, difficulty in diagnosis and lack of effective treatment. There are still many challenges in the prevention and treatment of pancreatic cancer, including the lack of early screening technology, the lack of public awareness of pancreatic cancer and uneven resource allocation. These factors make the prevention and treatment of pancreatic cancer urgent and complex.
[0003] The main treatments for pancreatic cancer include surgery, radiotherapy, and chemotherapy. Surgery is the best choice for treating early pancreatic cancer, but since many patients are already in the advanced stage when diagnosed, the indications for surgery are limited; in addition, the risk of complications and recurrence after surgery is also high. Chemotherapy is the most commonly used treatment for pancreatic cancer patients, especially in advanced patients, which can effectively relieve symptoms and prolong survival, but its side effects such as nausea, vomiting, and decreased immune function cannot be ignored. Radiotherapy is usually used for adjuvant treatment or pain relief, and its efficacy is limited by the specific location and size of the tumor. In recent years, targeted therapy has gradually attracted attention, with the advantages of strong targeting and relatively small side effects. Targeted therapy can directly act on the specific molecular mechanisms of cancer cells and improve the accuracy of treatment, which is expected to not only improve the survival rate of patients, but also improve the quality of life. Therefore, among various treatment methods, targeted therapy provides new hope and ideas for the treatment of pancreatic cancer.
[0004] In recent years, the research on synthetic small molecule compounds in inducing cell apoptosis to treat pancreatic cancer has gradually become a hot topic. This type of small molecule compound can effectively induce apoptosis of cancer cells by targeting and regulating specific signaling pathways and molecular mechanisms in cells, thereby inhibiting the growth and spread of tumors. Studies have shown that a variety of small molecule compounds targeting Bcl-2 family proteins, p53 pathways and NF-kB signaling pathways have shown good anti-cancer activity. In addition, these small molecule compounds also have a certain degree of selectivity, which can kill cancer cells while minimizing damage to normal cells. However, in current clinical applications, small molecule compounds have the problem of low activity. Summary of the invention
[0005] In order to solve the above technical problems, the present invention provides a pyridine dimethoxyphenylamino urea derivative and a preparation method and application thereof.
[0006] To achieve the above objectives, the specific technical solutions of the present invention are as follows.
[0007] The first aspect of the present invention provides a pyridine bimethoxyphenylaminourea derivative, characterized in that the structural formula of the pyridine bimethoxyphenylaminourea derivative is as shown in formula (I):
[0008]
[0009] Wherein, R is any one or two substituents selected from halogen, C1-C6 alkyl, and methoxy.
[0010] The pyridine dimethoxyphenylaminourea derivatives of the present invention are small molecule compounds, have good activity, and inhibit the growth of pancreatic cancer cells by inducing cell apoptosis, thereby achieving the effect of inhibiting tumors.
[0011] In another preferred embodiment, the specific structural formula of the pyridine bimethoxyphenylaminourea derivative is as shown in Formula (1) to Formula (4):
[0012]
[0013]
[0014] The second aspect of the present invention provides a method for preparing a pyridine dimethoxyphenylaminourea derivative, comprising the following steps:
[0015] The 6-(4-methoxyphenyl)-2-methylnicotinylhydrazide represented by formula (5) and compound A are subjected to a condensation reaction at 70°C to 80°C in an organic solvent environment to obtain a pyridine dimethoxyphenylaminourea / thiourea derivative;
[0016] The molar ratio of compound A to substituted phenyl isocyanate / substituted phenyl thioisocyanate is 1.0-1.1:1;
[0017] The compound A is a substituted phenyl isocyanate or a substituted phenyl thioisocyanate;
[0018]
[0019] In another preferred embodiment, the condensation reaction time is 8h to 12h; the organic solvent is anhydrous ethanol;
[0020] The substituted phenyl isocyanate is any one of toluene isocyanate, 2-chlorophenyl isocyanate, and 4-methoxyphenyl isocyanate;
[0021] The substituted phenylthioisocyanate is 1,2-dichloro-4-phenylisothiocyanate.
[0022] In another preferred embodiment, the specific preparation method of the 6-(4-methoxyphenyl)-2-methylnicotinic acid hydrazide is as follows:
[0023] Mix 6-(4-methoxyphenyl)-2-methylnicotinic acid ethyl ester represented by formula (6), hydrazine hydrate and anhydrous ethanol, react at 120° C. to 130° C. for 8 h to 12 h, filter and dry to obtain 6-(4-methoxyphenyl)-2-methylnicotinic acid hydrazide;
[0024] The molar ratio of 6-(4-methoxyphenyl)-2-methylnicotinate to hydrazine hydrate is 1:8.2-8.5;
[0025] The volume ratio of hydrazine hydrate to anhydrous ethanol is 8.5 to 9:10;
[0026]
[0027] In another preferred embodiment, the specific preparation method of the 6-(4-methoxyphenyl)-2-methylnicotinate is as follows:
[0028] 3-dimethylamino-1-methoxyphenylpropiophenone, ethyl acetoacetate, ammonium acetate and glacial acetic acid represented by formula (7) are mixed, reacted at 125° C. to 130° C. for 8 h to 12 h, and purified to obtain 6-(4-methoxyphenyl)-2-methylnicotinate ethyl ester;
[0029] The molar ratio of 3-dimethylamino-1-methoxypropiophenone, ethyl acetoacetate and ammonium acetate is 1:1-1.1:7-7.5;
[0030] The mass ratio of the glacial acetic acid to 3-dimethylamino-1-methoxypropiophenone is 10 to 20:1;
[0031]
[0032] In another preferred embodiment, the specific process of the purification is as follows:
[0033] The mixture was extracted with ethyl acetate, the extract was dried, filtered, rotary evaporated, and sanded to obtain a crude product; the crude product was eluted with a silica gel column to obtain 6-(4-methoxyphenyl)-2-methylnicotinate ethyl ester;
[0034] The eluent of the silica gel column is ethyl acetate and petroleum ether;
[0035] The volume ratio of the ethyl acetate to the petroleum ether is 1:3.5-4.
[0036] In another preferred embodiment, the specific preparation process of the 3-dimethylamino-1-methoxypropiophenone is as follows:
[0037] Methoxyacetophenone and N,N-dimethylformamide dimethyl acetal are mixed, kept at 110°C to 120°C for 45h to 48h, filtered, washed with ethyl acetate, and dried to obtain 3-dimethylamino-1-methoxypropiophenone;
[0038] The molar ratio of the methoxyacetophenone to the N,N-dimethylformamide is 1:3-3.5.
[0039] The third aspect of the present invention provides a use of a pyridine dimethoxyphenylaminourea derivative in the preparation of a drug for treating pancreatic cancer.
[0040] Compared with the prior art, the present invention has the following beneficial effects:
[0041] (1) The pyridine dimethoxyphenylaminourea derivatives of the present invention have good activity and inhibit the growth of pancreatic cancer cells by inducing cell apoptosis, thereby achieving the effect of inhibiting tumors. The IC values of the pyridine dimethoxyphenylaminourea derivatives of the present invention on five different pancreatic cancer cell lines, BxPC-3, AsPC-1, PANC-1, and MIAPaCa-2 were 50 The lowest values were 4.39 μM, 1.52 μM, 6.83 μM and 3.68 μM, respectively. This indicates that the pyridine dimethoxyphenylamino urea derivatives of the present invention have good inhibitory effects on different pancreatic cancer cells.
[0042] (2) The preparation process of the pyrimidine-linked methoxyphenylamino urea derivatives in the present invention utilizes a molecular hybridization strategy to design a new small molecule compound, and the entire preparation process is simple, suitable for large-scale production, and effectively reduces the synthesis cost. BRIEF DESCRIPTION OF THE DRAWINGS
[0043] Figure 1 The invention is a synthetic route of pyridine dimethoxyphenylaminourea derivatives, wherein a is N,N-dimethylformamide dimethyl acetal, b is ethyl acetoacetate, ammonium acetate and acetic acid, c is hydrazine hydrate with a mass percentage of 80%, and d is ethanol and iso(thio)cyanate.
[0044] Figure 2 The figure shows that H-23 inhibits the proliferation of pancreatic cancer cells; A is a staining diagram of the H-23 cell clone formation experiment after treatment with H-23 and DMSO, and B is a quantitative diagram of the number of H-23 clones; DMSO is dimethyl sulfoxide.
[0045] Figure 3The figure is a diagram of H-23-induced apoptosis of MIAPaCa-2 cells; wherein, A is a diagram showing the effect of H-23 on cleaved PARP protein, B is a quantitative diagram showing the relative expression of cleaved PARP protein, and C is a flow cytometry detection diagram of MIAPaCa-2 cells treated with different concentrations of H-23; DMSO is dimethyl sulfoxide. DETAILED DESCRIPTION
[0046] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.
[0047] Unless otherwise specified, the experimental methods described in the following examples are all conventional methods; the reagents and materials described are all commercially available unless otherwise specified.
[0048] The research on small molecule compounds in inducing cell apoptosis to treat pancreatic cancer has gradually become a hot topic. However, there are still many challenges in clinical application, such as low drug activity. In addition, there are many technical challenges in the treatment of pancreatic cancer, mainly reflected in the preparation process, such as cumbersome synthesis methods and high costs. These factors not only increase the economic burden of drug research and development and production, but also limit its market application.
[0049] The embodiments of the present invention provide a pyridine dimethoxyphenylamino urea derivative and a preparation method and application thereof. The pyridine dimethoxyphenylamino urea derivative has good activity and inhibits the growth of pancreatic cancer cells by inducing cell apoptosis, thereby achieving the effect of inhibiting tumors. In addition, by optimizing the synthesis route and preparation process of small molecule compounds, the production cost can be effectively reduced, the synthesis efficiency can be improved, and larger-scale production can be achieved. This will help to improve the accessibility of drugs in clinical applications, provide more effective and economical treatment options for pancreatic cancer patients, and demonstrate the broad development potential in this field. The preparation process of the pyridine dimethoxyphenylamino urea derivative in the present invention is as follows: Figure 1 shown.
[0050] The general structural formula of the pyridine dimethoxyphenylaminourea derivatives in the following embodiments is shown in formula (I):
[0051]
[0052] The R is any one or two substituents selected from halogen, C1-C6 alkyl, and methoxy.
[0053] The following is a detailed description of the preparation method of a pyridine dimethoxyphenylamino urea derivative.
[0054] Pancreatic cancer cells BxPC-3, AsPC-1, PANC-1, and MIAPaCa-2 were purchased from Wuhan Pronocell Life Science Technology Co., Ltd.
[0055] The specific preparation process of 6-(4-methoxyphenyl)-2-methylnicotinate in the following example is as follows:
[0056] S1. Add 0.04 mol of p-methoxyacetophenone and 16.0 mL of N,N-dimethylformamide dimethyl acetal (0.12 mol) into a dry 50 mL round-bottom flask, heat to 110 °C in an oil bath with stirring, keep warm for 48 h, monitor by TLC, and cool to room temperature after the reaction is complete. A yellow solid precipitates, filter it, wash it with ethyl acetate, and dry it to obtain 6.6 g of yellow solid 3-dimethylamino-1-methoxypropiophenone with a yield of 81% and a melting point of 95.2-95.6 °C.
[0057] The structure of 3-dimethylamino-1-methoxypropiophenone is shown in formula (7):
[0058]
[0059] Popper data is: 1 13C NMR(CDCl3): δ187.4,161.9,153.8,133.2,129.4,113.3,91.7,55.3,45.0,37.0; HRMS(ES): m / z calcdfor C 12 H 16 NO2206.1176, found 206.1189.
[0060] S2. In a dry 50mL round-bottom flask, 1.56g of 3-dimethylamino-1-methoxypropiophenone (7.6mmol), 1.07mL of ethyl acetoacetate (8.4mmol), 4.71g of ammonium acetate (54mmol), and 15mL of glacial acetic acid (15.735g) were added in sequence, and the mixture was heated to 125°C in a stirred oil bath, and refluxed for 8h. The reaction was monitored by TLC tracking. After the reaction was completed, the mixture was cooled to room temperature, extracted three times with 50mL of ethyl acetate, and eluted with ethyl acetate and petroleum ether in a volume ratio of 1:4 through silica gel column chromatography to obtain 1.73g of 6-(4-methoxyphenyl)-2-methylnicotinate as a light yellow solid with a yield of 84% and a melting point of 62.2-63.5g.
[0061] The structure of 6-(4-methoxyphenyl)-2-methylnicotinate is shown in formula (6):
[0062]
[0063] Pope data: 1 H NMR(600MHz,DMSO-d6):d 8.16(d,J=8.1Hz,1H),8.10(d,J=8.8Hz,2H),7.81(d,J=8.4Hz,1H),7.04(d,J=8. 8Hz, 2H), 4.30 (q, J = 7.2Hz, 2H), 3.82 (s, 3H), 2.76 (s, 3H), 1.32 (t, J = 7.2Hz, 3H); 13 C NMR (150MHz, DMSO-d6): d 166.3, 161.3, 159.0, 157.9, 139.6, 130.3, 128.9, 122.9, 116.9, 114.6, 61.2, 55.7, 25.3, 14.5.
[0064] S3, 4.54g of 6-(4-methoxyphenyl)-2-methylnicotinic acid ethyl ester (15.75mmol), 8.55mL of 80% by mass hydrazine hydrate and 10mL of anhydrous ethanol were added to a dry 25mL round-bottom flask, stirred and heated to 120℃ reflux, kept warm for 12h, and monitored by TCL tracking. After the reaction was complete, it was cooled to room temperature, and solid was generated. It was filtered, washed and dried to obtain 3.42g of light yellow solid 6-(4-methoxyphenyl)-2-methylnicotinic acid hydrazide, with a yield of 80%, and mp of 198.9~199.8℃.
[0065] The structure of 6-(4-methoxyphenyl)-2-methylnicotinic acid hydrazide is shown in formula (5):
[0066]
[0067] Spectrum data: 1H NMR (600MHz, DMSO-d6): d 9.60 (s, 1H), 8.26-7.95 (m, 2H), 7.88-7.56 (m, 2H), 7.30-6.79 (m, 2H), 4.55 (s, 2H), 3.82 (d, J=2.2Hz, 3H), 2.60 (s, 3H); 13C NMR (150MHz, DMSO-d6): d 167.8, 160.8, 156.0, 155.9, 136.8, 130.9, 128.7, 128.5, 116.5, 114.6, 55.7, 23.5; HRMS (+): calcd for C14H16N3O2 [M+H] + 258.1237, found 258.122.
[0068] Example 1
[0069] A method for preparing a pyridine-dimethoxyphenylaminourea derivative comprises the following steps:
[0070] In a dry reaction bottle, add 0.43mmol of p-toluene isocyanate, 5mL of ethanol, and 0.1g of 6-(4-methoxyphenyl)-2-methylnicotinylhydrazide (0.39mmol) in sequence, heat to 70°C under stirring, and react for 8h. The reaction is complete when detected by thin layer chromatography, and the reaction is stopped. The reaction solution is cooled to room temperature, filtered, the filter cake is washed with ethanol, and dried to obtain a white solid product 1-(2-(4-methoxyphenyl)-6-methylnicotinyl)-4-p-tolyl semicarbazide with a yield of 86%. The melting point is 217.6-218.2°C.
[0071] The structural formula of the 1-(2-(4-methoxyphenyl)-6-methylnicotinoyl)-4-p-tolyl semicarbazide is shown in formula (1):
[0072]
[0073] Pope data: 1 H NMR(600MHz, DMSO-d6):10.14(d,J=1.8Hz,1H),8.81(s,1H),8.21(d,J=1.7Hz,1H),8.14-8.07(m,2H),7.90(d,J=7.9Hz,1H), 7.83(d,J=8.1Hz,1H),7.38(d,J=8.4Hz,2H),7.09(d,J=8.3Hz,2H),7.07-7.05(m,2H),3.83(s,3H),2.67(s,3H),2.24(s,3H); 13C NMR(150MHz,DMSO-d6):168.5,161.0,156.5,156.3,155.9,137.5,137.1, 131.2,130.9,129.6,128.6,127.9,119.0,116.5,114.6,55.7,23.6,20.8.
[0074] ESI-HRMS(+):m / z calcd for C 22 H 23 N4O3 + [M+H] + 391.1770, found 391.1762.
[0075] Example 2
[0076] A method for preparing a pyridine-dimethoxyphenylaminourea derivative comprises the following steps:
[0077] In a 25mL round-bottom flask, add 0.1g of 6-(4-methoxyphenyl)-2-methylnicotinylhydrazide (0.39mmol), 5mL of anhydrous ethanol, and 0.42mmol of 2-chlorophenylisocyanate, stir and heat to 80℃ and reflux, react for 8h, TLC detection shows that the reaction is over, and the reaction is stopped. Filter and wash with ethanol to obtain a white solid product 4-(2-chlorophenyl)-1-(2-(4-methoxyphenyl)-6-methylnicotinyl) semicarbazide with a yield of 78% and a melting point of 216.4-217.1℃;
[0078] The structural formula of the 4-(2-chlorophenyl)-1-(2-(4-methoxyphenyl)-6-methylnicotinoyl) semicarbazide is shown in formula (2):
[0079]
[0080] Pope data: 1 H NMR(600MHz,DMSO-d6):10.31(s,1H),9.01(s,1H),8.38(s,1H),8.17-8.04(m,3H),7.91-7.76(m ,2H),7.48(d,J=7.7Hz,1H),7.31(t,J=7.5Hz,1H),7.10-6.92(m,3H),3.83(s,3H),2.66(s,3H).
[0081] 13C NMR(150MHz,DMSO-d6):168.4,161.0,156.6,156.2,155.4,137.1,136.2,130.8,129.7,12 8.7,128.1,127.7,124.0,116.6,114.6,114.6,114.6,100.0,55.7,23.5; ESI-HRMS(+):m / z calcd for C 21 H 20 ClN4O3 + [M+H] + 411.1218, found 411.1210.
[0082] Example 3
[0083] A method for preparing a pyridine-dimethoxyphenylaminourea derivative comprises the following steps:
[0084] In a 25mL round-bottom flask, add 0.1g of 6-(4-methoxyphenyl)-2-methylnicotinylhydrazide (0.39mmol), 5mL of anhydrous ethanol, and 0.42mmol of 4-methoxyphenylisocyanate, stir and heat to 80℃ and reflux, react for 10h, and stop the reaction after TLC detection. Filter and wash with ethanol to obtain a white solid product 4-(4-methoxyphenyl)-1-(2-(4-methoxyphenyl)-6-methylnicotinyl) semicarbazide with a yield of 76% and a melting point of 259.4-260.0℃;
[0085] The structural formula of the 4-(4-methoxyphenyl)-1-(2-(4-methoxyphenyl)-6-methylnicotinoyl) semicarbazide is shown in formula (3):
[0086]
[0087] Pope data: 1 H NMR (600MHz, DMSO-d6): 10.13(s,1H),8.73(s,1H),8.18(s,1H),8.11(d,J=8.8Hz,2H),7.91(d,J=7.7Hz,1H),7.83(d,J =8.1Hz, 1H), 7.40 (d, J = 8.8Hz, 2H), 7.06 (d, J = 8.8Hz, 2H), 6.87 (d, J = 8.8Hz, 2H), 3.83 (s, 3H), 3.72 (s, 3H), 2.66 (s, 3H).
[0088] 13C NMR(150MHz,DMSO-d6):168.5,161.0,156.4,156.3,156.1,155.0,137.2,133.1,13 0.8,128.6,127.9,120.8,116.5,114.6,114.3,55.7,55.6,23.6; ESI-HRMS(+):m / z calcd for C 22 H 23 N4O4 + [M+H] + 407.1714, found 407.1715.
[0089] Example 4
[0090] A method for preparing a pyridine-dimethoxyphenylaminourea derivative comprises the following steps:
[0091] In a 25ml round-bottom flask, add 0.1g of 6-(4-methoxyphenyl)-2-methylnicotinylhydrazide (0.39mmol), 5mL of anhydrous ethanol, and 0.42mmol of 4-methylphenyl isothiocyanate, stir and heat to 80℃ and reflux, react for 12h, and stop the reaction after TLC detection. Filter and wash with ethanol to obtain a white solid product 1-(2-(4-methoxyphenyl)-6-methylnicotinyl)-4-p-tolylthiosemicarbazide, with a yield of 84% and a melting point of 185.2-185.6℃;
[0092] The structural formula of the 1-(2-(4-methoxyphenyl)-6-methylnicotinoyl)-4-p-tolylthiosemicarbazide is shown in formula (1):
[0093]
[0094] Pope data: 1 H NMR (600MHz, DMSO-d6): 10.42 (s, 1H), 9.72 (s, 2H), 8.12 (d, J = 8.4Hz, 2H), 7.85 (d, J = 8.1Hz, 1H), 7.33(s,2H),7.17(d,J=8.1Hz,2H),7.07(d,J=8.8Hz,2H),3.83(s,3H),2.69(s,3H),2.30(s,3H).
[0095] 13C NMR(150MHz,DMSO-d6):161.0,156.5,153.2,137.6,137.0,134.8,130.8,129.0,12 8.7,124.8,118.1,116.3,114.6,113.6,100.6,55.7,23.9,21.0; ESI-HRMS(+):m / z calcd for C 22 H 23 N4O2S + [M+H] + 407.1536, found 407.1536.
[0096] Example 5
[0097] A method for preparing a pyridine-dimethoxyphenylaminourea derivative comprises the following steps:
[0098] In a 25ml round-bottom flask, add 0.1g of 6-(4-methoxyphenyl)-2-methylnicotinylhydrazide (0.39mmol), 5mL of anhydrous ethanol, and 0.42mmol of 1,2-dichloro-4-phenylisothiocyanate, stir and heat to 80℃ and reflux, react for 8 hours, and the reaction is stopped after TLC detection. Filter and wash with ethanol to obtain a white solid product 4-(3,4-dichlorophenyl)-1-(2-(4-methoxyphenyl)-6-methylnicotinyl)thiosemicarbazide, with a yield of 85% and a melting point of 179.9-180.6℃;
[0099] The structural formula of the 4-(3,4-dichlorophenyl)-1-(2-(4-methoxyphenyl)-6-methylnicotinoyl)thiosemicarbazide is shown in formula (4):
[0100]
[0101] Pope data: 1 H NMR(600MHz, DMSO-d6):10.45(s,1H),10.03(s,1H),9.91(s,1H),8.12(d,J=8.8Hz,3H),7.87(d,J=8.1 Hz,2H),7.62(d,J=8.8Hz,1H),7.54(d,J=8.1Hz,1H),7.07(d,J=8.8Hz,2H),3.83(s,3H),2.68(s,3H).
[0102] 13C NMR(150MHz,DMSO-d6):168.0,161.0,156.7,139.9,137.6,130.8,130.3,130.2,12 8.7,128.5,127.6,127.5,126.7,126.2,116.3,114.6,55.7,24.1; ESI-HRMS(+):m / z calcd for C21H19Cl2N4O2S+[M+H]+461.0600,found461.059.
[0103] The 1-(2-(4-methoxyphenyl)-6-methylnicotinoyl)-4-p-tolyl semicarbazide in Example 1 was recorded as H-6; the 4-(2-chlorophenyl)-1-(2-(4-methoxyphenyl)-6-methylnicotinoyl) semicarbazide in Example 2 was recorded as H-11; the 4-(4-methoxyphenyl)-1-(2-(4-methoxyphenyl)-6-methylnicotinoyl) semicarbazide in Example 3 was recorded as H-14; the 1-(2-(4-methoxyphenyl)-6-methylnicotinoyl)-4-p-tolylthiosemicarbazide in Example 4 was recorded as H-17; and the 4-(3,4-dichlorophenyl)-1-(2-(4-methoxyphenyl)-6-methylnicotinoyl)thiosemicarbazide in Example 5 was recorded as H-23; and the following experiments were performed respectively.
[0104] 1. Toxicity test
[0105] The CCK-8 method was used to perform in vitro toxicity tests on the five compounds H-6, H-11, H-14, H-17 and H-23 in Examples 1 to 5, with gemcitabine as a positive control. The experiment set up six concentration gradients of 1.5625μM, 3.1255μM, 6.2555μM, 12.555μM, 25.5μM and 50.5μM. The OD value was measured after 48 hours of treatment, and the IC 50 The results are shown in Table 1.
[0106] Table 1 Half inhibitory concentration of compounds on cancer cells
[0107]
[0108]
[0109] From the results in Table 1, it can be seen that the five compounds in the examples of the present invention all showed certain inhibitory effects on different pancreatic cancer cell lines, namely BxPC-3, AsPC-1, PANC-1, and MIAPaCa-2, among which compound H-23 showed the best activity, which was equivalent to the activity of the positive drugs. Therefore, H-23 was selected as the active compound for further study.
[0110] 2. Effect of H-23 on cell clone formation ability
[0111] AsPC-1 and MIAPaCa-2 cells were treated with 2 μM DMSO and 2 μM H-23, respectively. The drug solution was changed every 3 days for 2 consecutive weeks. The results are shown in Figure 2 As shown, from Figure 2 It can be seen that H-23 significantly reduced the clone numbers of AsPC-1 and MIAPaCa-2 cells, further demonstrating the inhibitory effect of H-23 on the proliferation of pancreatic cancer cells.
[0112] Apoptosis plays a key role in anti-pancreatic cancer treatment. By inducing programmed cell death in cancer cells, tumor growth can be effectively inhibited. The ability of compound H-23 to induce apoptosis in pancreatic cancer cells was further tested. MIAPaCa-2 cells were treated with 0-, 1-, 2-, and 4-directed H-23, and then apoptosis was detected by Western Blot and flow cytometry. The results are shown in Figure 2. Figure 3 As shown. A is a Western Blot test of the effect of H-23 on cleaved PARP protein, GAPDH is used as an internal reference protein, B is a quantitative test of the relative expression of cleaved PARP protein in Figure A, and C is a flow cytometry test of the ability of different concentrations of H-23 to induce cell apoptosis. Figure 3 It can be seen that H-23 significantly induces cell apoptosis. H-23 may inhibit the growth of pancreatic cancer cells by inducing cell apoptosis, thereby achieving the effect of inhibiting tumors.
[0113] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.
[0114] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.
Claims
1. A pyridine bimethoxyphenylamino urea derivative, characterized in that: The structural formula of the pyridine bimethoxyphenylaminourea derivative is shown in formula (I): Wherein, R is any one or two substituents selected from halogen, C1-C6 alkyl, and methoxy.
2. The pyridine bimethoxyphenylaminourea derivative according to claim 1, characterized in that The specific structural formula of the pyridine bimethoxyphenylaminourea derivative is shown in formula (1) to formula (4):
3. A method for preparing the pyridine bimethoxyphenylaminourea derivative according to claim 1, characterized in that: The following steps are involved: The 6-(4-methoxyphenyl)-2-methylnicotinylhydrazide represented by formula (5) and compound A are subjected to a condensation reaction at 70°C to 80°C in an organic solvent environment to obtain a pyridine dimethoxyphenylaminourea / thiourea derivative; The molar ratio of compound A to 6-(4-methoxyphenyl)-2-methylnicotinic acid hydrazide is 1.1 to 1:1; The compound A is a substituted phenyl isocyanate or a substituted phenyl thioisocyanate; 4. The method for preparing the pyridine bimethoxyphenylaminourea derivative according to claim 3, characterized in that: The condensation reaction time is 8h to 12h; The organic solvent is anhydrous ethanol; The substituted phenyl isocyanate is any one of toluene isocyanate, 2-chlorophenyl isocyanate, and 4-methoxyphenyl isocyanate; The substituted phenylthioisocyanate is 1,2-dichloro-4-phenylisothiocyanate.
5. The method for preparing the pyridine bimethoxyphenylaminourea derivative according to claim 3, characterized in that: The specific preparation method of the 6-(4-methoxyphenyl)-2-methylnicotinic acid hydrazide is as follows: Mix 6-(4-methoxyphenyl)-2-methylnicotinic acid ethyl ester represented by formula (6), hydrazine hydrate and anhydrous ethanol, react at 120° C. to 130° C. for 8 h to 12 h, filter and dry to obtain 6-(4-methoxyphenyl)-2-methylnicotinic acid hydrazide; The molar ratio of 6-(4-methoxyphenyl)-2-methylnicotinate to hydrazine hydrate is 1:8.2-8.5; The volume ratio of hydrazine hydrate to anhydrous ethanol is 8.5 to 9:10; 6. The method for preparing the pyridine bimethoxyphenylaminourea derivative according to claim 5, characterized in that: The specific preparation method of the 6-(4-methoxyphenyl)-2-methylnicotinate is as follows: 3-dimethylamino-1-methoxyphenylpropiophenone, ethyl acetoacetate, ammonium acetate and glacial acetic acid represented by formula (7) are mixed, reacted at 125° C. to 130° C. for 8 h to 12 h, and purified to obtain 6-(4-methoxyphenyl)-2-methylnicotinate ethyl ester; The molar ratio of 3-dimethylamino-1-methoxypropiophenone, ethyl acetoacetate and ammonium acetate is 1:1-1.1:7-7.5; The mass ratio of glacial acetic acid to 3-dimethylamino-1-methoxypropiophenone is 10 to 20:1; 7. The method for preparing the pyridine-bimethoxyphenylaminourea derivative according to claim 6, characterized in that: The specific process of the purification is as follows: The mixture was extracted with ethyl acetate, the extract was dried, filtered, and rotary evaporated to obtain a crude product; the crude product was eluted through a silica gel column to obtain 6-(4-methoxyphenyl)-2-methylnicotinate ethyl ester; The eluent of the silica gel column is ethyl acetate and petroleum ether; The volume ratio of ethyl acetate to petroleum ether is 1:3.5-4.
8. The method for preparing the pyridine-bimethoxyphenylaminourea derivative according to claim 5, characterized in that: The specific preparation process of the 3-dimethylamino-1-methoxypropiophenone is as follows: Methoxyacetophenone and N,N-dimethylformamide dimethyl acetal are mixed, kept at 110°C to 120°C for 45h to 48h, filtered, washed with ethyl acetate, and dried to obtain 3-dimethylamino-1-methoxypropiophenone; The molar ratio of the methoxyacetophenone to the N,N-dimethylformamide dimethyl acetal is 1:3-3.
5.
9. Use of the pyridine bimethoxyphenylaminourea derivative according to claim 1 in the preparation of a drug for treating pancreatic cancer.