A flavonoid derivative and its preparation method and application

Abstract

The present invention discloses a flavonoid derivative and a preparation method and application thereof, relating to the technical field of pharmaceutical chemistry. The flavonoid derivative comprises a chrysin derivative or a baicalein derivative. The general structural formula of the chrysin derivative is shown in Formula I-1, and the general structural formula of the baicalein derivative is shown in Formula I-2. The results show that the compound 2-phenyl-5-hydroxy-7-((6-(4-methoxypiperidine-1)-2-methylpyrimidin-4-yl))oxy)-4H-chromene-4-one (BY6) has significant cytotoxic activity (IC) against A549 cell lines. 50 :14.70μM), with higher inhibitory activity than cisplatin; and the best cytotoxic activity against PC‑3 cell line (IC 50 :9.18μM), and its inhibitory activity was comparable to that of cisplatin; its inhibitory activity against HCT116 cell line was comparable to that of cisplatin (IC 50 :19.47 μM).

Description

Technical Field

[0001] The present invention relates to the technical field of medicinal chemistry, and in particular to a flavonoid compound derivative and a preparation method and application thereof. Background Art

[0002] Flavonoids are a general term for a series of compounds composed of two benzene rings linked by three carbon atoms, i.e., compounds with a C6-C3-C6 structure. They are widely found in plants in nature and are secondary metabolites. Most flavonoids exist in plants as glycosides or carbonyl groups, bound to sugars. Some also exist in a free form.

[0003] Chrysin and baicalein are two naturally derived flavonoids with anti-inflammatory, antibacterial, antiviral, antimutagenic, and anticancer properties that are closely related to the inhibition of tumor occurrence and progression. Chrysin and baicalein have also attracted much attention in the development of anticancer drugs. Furthermore, these flavonoids are widely distributed in plants and have low toxicity, making them a very important resource in new drug development research. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the present invention aims to provide a flavonoid derivative and a preparation method and application thereof, aiming to develop a new drug with good anti-cancer activity.

[0005] The first aspect of the present invention is to provide a flavonoid derivative, wherein the flavonoid derivative includes a chrysin derivative or a baicalein derivative. The general structural formula of the chrysin derivative is shown in Formula I-1, and the general structural formula of the baicalein derivative is shown in Formula I-2:

[0006]

[0007] in,

[0008] in,

[0009] Compared with the prior art, the beneficial effects of the present invention are as follows: an intermediate is generated by reacting a flavonoid compound with 4,6-dichloro-2-methylpyrimidine, wherein the flavonoid compound includes chrysin or baicalein; the intermediate is reacted with a corresponding fatty amine to obtain 12 flavonoid compound derivatives, all of which are new compounds. Screening results of cancer cell proliferation inhibitory activity show that one of the compounds, BY6, has a cytotoxic activity against A549 cell lines higher than that of cisplatin; it exhibits the strongest inhibitory effect on PC-3 cell lines, with a cytotoxic activity equivalent to that of cisplatin; and its proliferation inhibitory activity against HCT116 is equivalent to that of cisplatin.

[0010] The second aspect of the present invention is to provide a method for preparing a flavonoid derivative, wherein the method is used to prepare the above-mentioned flavonoid derivative, and the method comprises:

[0011] reacting a flavonoid compound with 4,6-dichloro-2-methylpyrimidine to generate an intermediate, wherein the flavonoid compound includes chrysin or baicalein;

[0012] The intermediate is reacted with a corresponding fatty amine to obtain a flavonoid derivative.

[0013] According to one aspect of the above technical solution, when the flavonoid compound is chrysin, the synthesis route is as follows:

[0014]

[0015] Among them, fatty amine

[0016] Dissolving chrysin and 4,6-dichloro-2-methylpyrimidine in an organic solvent DMF at a first predetermined molar ratio, and stirring the mixture under the action of an alkaline reagent K2CO3 to obtain a chrysin intermediate;

[0017] The chrysin intermediate and the corresponding fatty amine 1 are dissolved in a solvent ethanol at a second preset molar ratio, and the temperature is raised to a preset temperature and reacted for a preset time under the action of an alkaline reagent DIPEA to obtain a chrysin derivative.

[0018] According to one aspect of the above technical solution, when the flavonoid compound is baicalein, the synthesis route is as follows:

[0019]

[0020] Among them, fatty amine

[0021] dissolving baicalein and 4,6-dichloro-2-methylpyrimidine in an organic solvent DMF at a first preset molar ratio, and stirring and reacting them under the action of an alkaline reagent K2CO3 to obtain a baicalein intermediate;

[0022] The baicalein intermediate and the corresponding fatty amine 2 are dissolved in a solvent ethanol at a second preset molar ratio, and the temperature is raised to a preset temperature and reacted for a preset time under the action of an alkaline reagent DIPEA to obtain a baicalein derivative.

[0023] According to one aspect of the above technical solution, the first preset molar ratio is 1:(0.7-1.3), and the second preset molar ratio is 1:(0.9-1.5).

[0024] According to one aspect of the above technical solution, the preset temperature is 70°C-90°C, and the preset time is 9h-12h.

[0025] According to one aspect of the above technical solution, the molar amount of the alkaline reagent K2CO3 is 10mmol-15mmol, the volume of the organic solvent DMF is 10mL-50mL, the molar amount of the alkaline reagent DIPEA is 1mmol-1.5mmol, and the volume of the ethanol is 1mL-10mL.

[0026] According to one aspect of the above technical solution, after the step of stirring the reaction under the action of the alkaline reagent K2CO3, the method further comprises:

[0027] Deionized water is added and stirred for 5-10 minutes, the filtrate is removed by filtration, and the filter cake is subjected to column chromatography to obtain a chrysin intermediate or a baicalein intermediate.

[0028] According to one aspect of the above technical solution, after the step of heating to a preset temperature and reacting for a preset time under the action of the alkaline reagent DIPEA, the method further comprises:

[0029] After the reaction is completed, the mixture is concentrated in vacuo, cooled to room temperature, washed with ethyl acetate or petroleum ether and recrystallized to obtain a chrysin derivative or a baicalein derivative.

[0030] The third aspect of the present invention is to provide the use of the flavonoid derivatives described above in the field of anti-cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the following description of the embodiments with reference to the accompanying drawings, in which:

[0032] Figure 1 is the general structural formula of the flavonoid derivatives of the present invention. DETAILED DESCRIPTION

[0033] To make the objectives, features, and advantages of the present invention more readily apparent, the following detailed description of specific embodiments of the present invention is provided in conjunction with the accompanying drawings. The accompanying drawings illustrate several embodiments of the present invention. However, the present invention may be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a more thorough and comprehensive understanding of the present invention.

[0034] It should be noted that when an element is referred to as being "fixed to" another element, it may be directly on the other element or there may be an element centered thereon. When an element is considered to be "connected to" another element, it may be directly connected to the other element or there may be an element centered thereon. The terms "vertical", "horizontal", "left", "right", "up", "down" and similar expressions used herein are for illustrative purposes only and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore should not be understood as limiting the present invention.

[0035] In the present invention, unless otherwise expressly specified or limited, the terms "installed," "connected," "connected," "fixed," and the like should be understood in a broad sense. For example, they may refer to fixed connections, detachable connections, or integral connections; they may refer to mechanical connections or electrical connections; they may refer to direct connections or indirect connections through an intermediate medium; and they may refer to internal communication between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on the specific circumstances. The term "and / or" as used herein includes any and all combinations of one or more of the relevant listed items.

[0036] The present invention provides a flavonoid derivative, wherein the flavonoid derivative includes a chrysin derivative or a baicalein derivative. The general structural formula of the chrysin derivative is shown in Formula I-1, and the general structural formula of the baicalein derivative is shown in Formula I-2:

[0037]

[0038] in,

[0039] in,

[0040] In addition, the present invention provides a method for preparing a flavonoid compound derivative, which is used to prepare the above-mentioned flavonoid compound derivative, and the preparation method comprises:

[0041] reacting a flavonoid compound with 4,6-dichloro-2-methylpyrimidine to generate an intermediate, wherein the flavonoid compound includes chrysin or baicalein;

[0042] The intermediate is reacted with a corresponding fatty amine to obtain a flavonoid derivative.

[0043] Specifically, when the flavonoid compound is chrysin, the synthesis route is as follows:

[0044]

[0045] Among them, fatty amine

[0046] Dissolving chrysin and 4,6-dichloro-2-methylpyrimidine in an organic solvent DMF at a first predetermined molar ratio, and stirring the mixture under the action of an alkaline reagent K2CO3 to obtain a chrysin intermediate;

[0047] The chrysin intermediate and the corresponding fatty amine 1 are dissolved in a solvent ethanol at a second preset molar ratio, and the temperature is raised to a preset temperature and reacted for a preset time under the action of an alkaline reagent DIPEA to obtain a chrysin derivative.

[0048] When the flavonoid compound is baicalein, the synthesis route is as follows:

[0049]

[0050] Among them, fatty amine

[0051] dissolving baicalein and 4,6-dichloro-2-methylpyrimidine in an organic solvent DMF at a first preset molar ratio, and stirring and reacting them under the action of an alkaline reagent K2CO3 to obtain a baicalein intermediate;

[0052] The baicalein intermediate and the corresponding fatty amine 2 are dissolved in a solvent ethanol at a second preset molar ratio, and the temperature is raised to a preset temperature and reacted for a preset time under the action of an alkaline reagent DIPEA to obtain a baicalein derivative.

[0053] It should be noted that 4,6-dichloro-2-methylpyrimidine at different sites reacts with chrysin and baicalein respectively under the action of an alkaline reagent to obtain a chrysin intermediate and a baicalein intermediate, respectively.

[0054] Preferably, the alkaline reagent is K2CO3 and the solvent is DMF.

[0055] Preferably, the first preset molar ratio is 1:(0.7-1.3), for example, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.2, or 1:1.3, but is not limited to the listed values, and other values ​​not listed within the numerical range are also applicable. The first preset molar ratio can effectively increase the yield and reduce the formation of by-products.

[0056] Preferably, the molar amount of the alkaline reagent KCO is 10 mmol to 15 mmol, for example, 10 mmol, 11 mmol, 12 mmol, 13 mmol, 14 mmol, or 15 mmol, but is not limited to the values ​​listed, and other values ​​not listed within the numerical range are also applicable. The alkaline reagent KCO can effectively catalyze the reaction, thereby increasing the reaction rate and yield.

[0057] Preferably, the volume of the organic solvent DMF is 10 mL-50 mL, for example, 10 mL, 20 mL, 30 mL, 40 mL or 50 mL, but is not limited to the listed values, and other values ​​not listed within the numerical range are also applicable.

[0058] Preferably, the preset temperature is 70°C-90°C, for example, 70°C, 75°C, 80°C, 85°C or 90°C, but is not limited to the listed values, and other unlisted values ​​within the numerical range are also applicable.

[0059] Preferably, the preset time is 9h-12h, for example, it can be 9h, 9.5h, 10h, 10.5h, 11h or 12h, but is not limited to the listed values, and other unlisted values ​​within the numerical range are also applicable.

[0060] After the step of stirring the reaction under the action of the alkaline reagent K2CO3, the method further comprises:

[0061] The reaction is stirred at room temperature overnight under the action of an alkaline reagent K2CO3. When thin layer chromatography (TLC) shows that the raw material chrysin or baicalein disappears, 80 mL-120 mL of deionized water is added and stirred for 5 min-10 min. The filtrate is removed by filtration, and the filter cake is subjected to column chromatography to obtain a chrysin intermediate or a baicalein intermediate.

[0062] The column chromatography was performed using petroleum ether solvent containing 6% ethyl acetate as the mobile phase for chromatography purification.

[0063] After the step of heating to a preset temperature and reacting for a preset time under the action of the alkaline reagent DIPEA, the method further comprises:

[0064] After the reaction is completed, the mixture is concentrated in vacuo, cooled to room temperature, washed with ethyl acetate or petroleum ether and recrystallized to obtain a chrysin derivative or a baicalein derivative.

[0065] In addition, the present invention also provides a use of flavonoid derivatives in the field of anti-cancer.

[0066] The present invention is further described below with specific examples:

[0067] Example 1

[0068] The first embodiment of the present invention provides a flavonoid derivative, wherein the flavonoid derivative (BY1) has a general structural formula as shown in Formula I-1-1:

[0069]

[0070] The specific preparation route is:

[0071]

[0072] 4,6-Dichloro-2-methylpyrimidine (1.79 g, 11.0 mmol), chrysin (2.54 g, 10.0 mmol), and K2CO3 (1.80 g, 13.0 mmol) were added to a 50 mL round-bottom flask, followed by the addition of DMF (20 mL) and stirring at room temperature overnight. After thin-layer chromatography (TLC) indicated the disappearance of the raw chrysin, water (100 mL) was added and stirred for 5 minutes. The mixture was filtered and the filter cake was purified by column chromatography (silica gel, petroleum ether containing 6% ethyl acetate as the mobile phase) to obtain the chrysin intermediate (3.6 g, 9.5 mmol, 95% yield).

[0073] fatty amines

[0074] The chrysin intermediate (0.38 g, 1 mmol), the corresponding fatty amine 1 (1.2 mmol), and N,N-diisopropylethylamine (DIPEA, 209 μL, 1.2 mmol) were dissolved in ethanol (2 mL) and heated with stirring at 80°C for 10 h. After completion of the reaction, the mixture was concentrated in vacuo, cooled to room temperature, and recrystallized from ethyl acetate / petroleum ether to obtain the chrysin derivative BY1 in a 72% yield.

[0075] The structural identification data is: 2-phenyl-5-hydroxy-7-((6-(2-methoxyethylamino)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (BY1): 1 H NMR (400MHz, CDCl3) δ12.76 (s, 1H), 7.99-7.80 (m, 2H), 7.63-7.46 (m, 3H), 6.78 (d, J=2.1Hz, 1H), 6.73 (s, 1H), 6 .58(d, J=2.1Hz, 1H), 5.70(s, 1H), 5.30(s, 1H), 3.62-3.53(m, 2H), 3.53-3.43(m, 2H), 3.38(s, 3H), 2.44(s, 3H); 13C NMR (101MHz, CDCl3) δ182.87, 168.87, 168.44, 165.19, 164.63, 162.15, 159.61, 157.30, 132.20, 131.13, 129. 25, 126.49, 108.17, 106.14, 104.35, 99.53, 95.10, 77.48, 77.16, 76.84, 70.83, 58.95, 41.28, 25.90; IR(KBr)ν max (cm -1 ): 3425, 3251, 3098, 2889, 1663, 1617, 1200, 1149, 1113, 993, 830, 682; ESI-MS displays a peak at m / z 420.0, calcd for [M+H] + :420.4445.

[0076] Example 2

[0077] The second embodiment of the present invention provides a flavonoid derivative (BY2) having a general structural formula as shown in Formula I-1-2:

[0078]

[0079] The specific preparation route is:

[0080]

[0081] The difference from Example 1 is that: fatty amine The chrysin derivative BY2 was generated in 90% yield.

[0082] The structural identification data is: 2-phenyl-5-hydroxy-7-((6-(4-isopropylpiperazinyl)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (BY2): 1 H NMR (400MHz, CDCl3) δ12.74 (s, 1H), 7.94-7.79 (m, 2H), 7.61-7.37 (m, 3H), 6.74 (d, J=2.0Hz, 1H), 6.69 (s, 1H), 6.55 (d, J =2.0Hz, 1H), 5.85 (s, 1H), 3.81-3.48 (m, 4H), 2.83-2.67 (m, 1H), 2.66-2.51 (m, 4H), 2.43 (s, 3H), 1.07 (d, J = 6.5Hz, 6H); 13C NMR (101MHz, CDCl3) δ182.85, 169.27, 168.12, 164.66, 164.58, 162.18, 159.97, 157.34, 132.14, 131.2 3, 129.24, 126.49, 108.06, 106.17, 104.15, 99.26, 85.16, 54.84, 48.37, 44.44, 26.16, 18.47; IR(KBr)ν max (cm -1 ): 3436, 2967, 2853, 1655, 1597, 1549, 1198, 1136, 853, 811, 687, 635; ESI-MS displays a peak at m / z 474.0, calcd for [M+H] + :473.5525.

[0083] Example 3

[0084] The third embodiment of the present invention provides a flavonoid derivative (BY3) having a general structural formula as shown in Formula I-1-3:

[0085]

[0086] The specific preparation route is:

[0087]

[0088] The difference from Example 1 is that: fatty amine The chrysin derivative BY3 was generated in 71% yield.

[0089] The structural identification data is: 2-phenyl-5-hydroxy-7-((6-(3-hydroxypropylamino)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (BY3): 1 H NMR (400MHz, CDCl3) δ12.78 (s, 1H), 7.97-7.83 (m, 2H), 7.64-7.38 (m, 3H), 6.77 (d, J=2.1Hz, 1H), 6.73 (s, 1H), 6.57 ( d, J=2.1Hz, 1H), 5.70 (s, 1H), 5.16 (s, 1H), 3.73-3.63 (m, 2H), 3.60-3.38 (m, 2H), 2.45 (s, 3H), 1.78 (t, J=6.1Hz, 2H); 13C NMR (101MHz, CDCl3) δ182.88, 168.73, 168.28, 165.39, 164.74, 163.69, 162.14, 159.42, 157.31, 132 .28, 131.04, 129.27, 126.50, 108.23, 106.11, 104.37, 99.64, 53.55, 29.79, 25.75, 25.72; IR(KBr)ν max (cm -1 ): 3421, 3069, 1657, 1616, 1200, 819, 677; ESI-MS displays peak at m / z 420.0, calcd for [M+H] + :420.4445.

[0090] Example 4

[0091] The fourth embodiment of the present invention provides a flavonoid derivative (BY4) having a general structural formula as shown in Formula I-1-4:

[0092]

[0093] The specific preparation route is:

[0094]

[0095] The difference from Example 1 is that: fatty amine The chrysin derivative BY4 was generated in 69% yield.

[0096] The structural identification data is: 2-phenyl-5-hydroxy-7-((6-(tetrahydropyrrol-1)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (BY4): 1 H NMR (400MHz, CDCl3) δ12.74 (s, 1H), 7.87 (d, J = 7.1Hz, 2H), 7.66-7.41 (m, 3H), 6.75 (d, J = 1.5Hz, 1H), 6.70 (s, 1H), 6.56 (d, J=1.5Hz, 1H), 5.64 (s, 1H), 3.95-2.93 (m, 4H), 2.47 (s, 3H), 2.25-1.56 (m, 4H); 13 C NMR (101 MHz, CDC l3)δ182.86, 168.21, 168.02, 164.57, 162.69, 162.18, 160.13, 157.37, 132.16, 131.21, 129.25, 126.49, 108.00, 106.15, 103.98, 99.10, 85.51, 46.81, 26.08, 25.34; IR(KBr)ν max (cm -1 ): 3062, 2967, 2926, 1658, 1609, 1344, 1192, 821, 767, 687, 674; ESI-MS displays a peak at m / z 417.0, calcd for [M+H] + :416.4565.

[0097] Example 5

[0098] The fifth embodiment of the present invention provides a flavonoid derivative (BY5) having a general structural formula as shown in Formula I-1-5:

[0099]

[0100] The specific preparation route is:

[0101]

[0102] The difference from Example 1 is that: fatty amine The chrysin derivative BY5 was generated in 77% yield.

[0103] The structural identification data is: 2-phenyl-5-hydroxy-7-((6-(N-methyl-2-hydroxyethylamino)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (BY5): 1 H NMR (400MHz, CDCl3) δ12.76 (s, 1H), 7.97-7.83 (m, 2H), 7.66-7.39 (m, 3H), 6.77 (d, J=2.1Hz, 1H), 6.73 (s, 1H), 6.58 (d, J=2.1Hz, 1H), 5.82 (s, 1H), 3.91-3.84 (m, 2H), 3.83-3.74 (m, 2H), 3.06 (s, 3H), 2.44 (s, 3H); 13C NMR (101MHz, CDCl3) δ182.88, 168.99, 167.70, 165.28, 164.63, 162.16, 159.68, 157.30, 132.21, 13 1.14, 129.26, 126.50, 108.16, 106.15, 104.25, 99.38, 85.18, 62.51, 53.60, 37.42, 25.91; IR(KBr)ν max (cm -1 ): 3411, 3076, 2925, 1664, 1608, 1452, 1199, 819, 677; ESI-MS displays apeak at m / z 420.0, calcd for [M+H] + :420.4445.

[0104] Example 6

[0105] The sixth embodiment of the present invention provides a flavonoid derivative (BY6) having a general structural formula as shown in Formula I-1-6:

[0106]

[0107] The specific preparation route is:

[0108]

[0109] The difference from Example 1 is that: fatty amine The chrysin derivative BY6 was generated in 85% yield.

[0110] The structural identification data is: 2-phenyl-5-hydroxy-7-((6-(4-methoxypiperidin-1)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (BY6): 1 H NMR (400MHz, CDCl3) δ12.76 (s, 1H), 7.93-7.85 (m, 2H), 7.61-7.48 (m, 3H), 6.76 (d, J=2.1Hz, 1H), 6.72 (s, 1H), 6.57 (d, J=2.1Hz , 1H), 5.89(s, 1H), 4.04-3.80(m, 2H), 3.53-3.44(m, 1H), 3.43-3.24(m, 5H), 2.45(s, 3H), 2.01-1.85(m, 2H), 1.71-1.52(m, 2H); 13C NMR (101MHz, CDCl3) δ182.87, 169.27, 168.14, 164.56, 164.46, 162.14, 160.00, 157.33, 132.16, 131.1 9, 129.25, 126.49, 108.01, 106.15, 104.06, 99.17, 85.14, 75.72, 55.88, 41.60, 30.32, 26.21; IR(KBr)ν max (cm -1 ): 3440, 3068, 2933, 1657, 1602, 1447, 1196, 824, 681; ESI-MS displays a peak at m / z 461.0, calcd for [M+H] + :460.5095.

[0111] Example 7

[0112] The seventh embodiment of the present invention provides a flavonoid derivative (BY7) having a general structural formula as shown in Formula I-1-7:

[0113]

[0114] The specific preparation route is:

[0115]

[0116] The difference from Example 1 is that: fatty amine The chrysin derivative BY7 was generated in 77% yield.

[0117] The structural identification data is: 2-phenyl-5-hydroxy-7-((6-(N-methyl-n-propylamino)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (BY7): 1 H NMR (400MHz, CDCl3) δ12.74 (s, 1H), 7.98-7.79 (m, 2H), 7.67-7.45 (m, 3H), 6.76 (d, J=2.1Hz, 1H), 6.72 (s, 1 H), 6.57 (d, J=2.1Hz, 1H), 5.77 (s, 1H), 3.05 (s, 3H), 2.45 (s, 3H), 1.70-1.53 ​​(m, 4H), 0.93 (t, J=7.4Hz, 3H); 13C NMR (101MHz, CDCl3) δ182.86, 168.68, 167.82, 164.59, 164.52, 162.12, 160.25, 157.35, 132.13, 131.2 3, 129.24, 126.48, 107.89, 106.14, 103.87, 98.96, 84.82, 51.34, 35.85, 26.23, 20.51, 11.45; IR(KBr)ν max (cm -1 ): 3447, 3084, 2965, 1660, 1603, 1227, 812, 671; ESI-MS displays a peak at m / z419.0, calcd for [M+H] + :418.4725.

[0118] Example 8

[0119] The eighth embodiment of the present invention provides a flavonoid derivative (BY8) having a general structural formula as shown in Formula I-1-8:

[0120]

[0121] The specific preparation route is:

[0122]

[0123] The difference from Example 1 is that: fatty amine The chrysin derivative BY8 was generated in 89% yield.

[0124] The structural identification data is: 2-phenyl-5-hydroxy-7-((6-(3,5-dimethylpiperidin-1)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (BY8): 1 H NMR (400MHz, CDCl3) δ12.73 (s, 1H), 7.87 (d, J=6.9Hz, 2H), 7.63-7.38 (m, 3H ), 6.74 (d, J = 1.6Hz, 1H), 6.70 (s, 1H), 6.55 (d, J = 1.6Hz, 1H), 5.89 (s, 1H), 4 .60-3.92(m, 2H), 2.44(s, 3H), 2.32(t, J=12.2Hz, 2H), 1.85(d, J=12.9Hz, 1 H), 1.72-1.55 (m, 2H), 0.94 (d, J=6.5Hz, 6H), 0.82 (dd, J=24.5, 12.1Hz, 1H); 13C NMR (101MHz, CDCl3) δ182.88, 169.13, 168.12, 164.55, 164.34, 162.17, 160.28, 157.39, 132.12, 131.3 1, 129.25, 126.51, 107.94, 106.19, 103.91, 98.99, 85.38, 51.60, 42.65, 31.04, 26.22, 19.27; IR(KBr)ν max (cm -1 ): 3435, 3083, 2953, 2924, 2844, 1661, 1601, 1447, 1209, 1143, 853, 811, 766, 671; ESI-MS displays a peak at m / z 459.0, calcd for [M+H] + :458.5375.

[0125] Example 9

[0126] The ninth embodiment of the present invention provides a flavonoid derivative (HQ1) having the general structural formula shown in Formula I-2-1:

[0127]

[0128] The specific preparation route is:

[0129]

[0130] 4,6-Dichloro-2-methylpyrimidine (1.79 g, 11.0 mmol), baicalein (2.70 g, 10.0 mmol), and K2CO3 (1.80 g, 13.0 mmol) were added to a 50 mL round-bottom flask, followed by the addition of DMF (20 mL) and stirring at room temperature overnight. After thin-layer chromatography (TLC) showed the disappearance of the raw baicalein, water (100 mL) was added and stirred for 5 minutes. The mixture was filtered, and the filter cake was purified by column chromatography (silica gel, petroleum ether containing 6% ethyl acetate as the mobile phase) to obtain the baicalein intermediate (3.33 g, 8.4 mmol, 84% yield).

[0131] fatty amines

[0132] The baicalein intermediate (0.4 g, 1 mmol), the corresponding fatty amine 2 (1.2 mmol), and N,N-diisopropylethylamine (DIPEA, 209 μL, 1.2 mmol) were dissolved in ethanol (2 mL) and heated with stirring at 80°C for 10 h. After completion of the reaction, the mixture was concentrated in vacuo, cooled to room temperature, and recrystallized from ethyl acetate / petroleum ether to obtain the baicalein derivative HQ1 in a 71% yield.

[0133] The structural identification data is: 2-phenyl-5,7-dihydroxy-6-((6-(3-hydroxypropylamino)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (HQ1): 1 H NMR (400MHz, DMSO-d6) δ8.20-7.88(m, 2H), 7.69-7.41(m, 3H), 6.82(s, 1H), 6.74(s, 1H) , 6.11 (s, 1H), 3.54-3.39 (m, 2H), 2.82 (t, J=7.0Hz, 2H), 2.43 (s, 3H), 1.75-1.54 (m, 2H); 13 C NMR (101MHz, DMSO-d6) δ180.30, 170.52, 168.24, 166.58, 161.42, 160.22, 155.52, 151.66, 131.38, 1 29.06, 126.15, 126.01, 125.82, 104.19, 103.14, 99.40, 96.12, 57.94, 36.74, 30.26, 25.29; IR(KBr)ν max (cm -1 ): 3416, 3069, 2926, 1668, 1607, 1578, 1446, 1181, 878, 827, 768, 578; ESI-MS displays a peak at m / z 437.0, calcd for [M+H] + :436.4435.

[0134] Example 10

[0135] The tenth embodiment of the present invention provides a flavonoid derivative (HQ2) having a general structural formula as shown in Formula I-2-2:

[0136]

[0137] The specific preparation route is:

[0138]

[0139] The difference from Example 9 is that: fatty amine The baicalein derivative HQ2 was generated in 73% yield.

[0140] The structural identification data is: 2-phenyl-5,7-dihydroxy-6-((6-(isopentylamino)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (HQ2): 1H NMR (400MHz, DMSO-d6) δ8.23-7.84(m, 2H), 7.68-7.41(m, 3H), 6.89(s, 1H), 6.80(s, 1H), 6.22(s, 1H), 2.85 -2.68 (m, 1H), 2.43 (s, 3H), 1.64-1.50 (m, 1H), 1.45-1.28 (m, 2H), 1.11-0.99 (m, 1H), 0.85 (d, J=6.6Hz, 6H) ; 13CNMR (101MHz, DMSO-d6) δ180.69, 170.35, 168.23, 164.95, 161.82, 160.25, 155.24, 151.77, 131.48, 13 1.22, 129.05, 126.08, 125.40, 104.36, 103.26, 95.81, 93.35, 37.27, 35.94, 25.28, 24.98, 22.12; IR(KBr)ν max (cm -1 ): 3428, 3071, 2956, 2927, 1654, 1556, 1448, 1388, 1178, 889, 771, 688, 583; ESI-MS displays a peak at m / z 449.0, calcd for [M+H] + :448.4985.

[0141] Example 11

[0142] The eleventh embodiment of the present invention provides a flavonoid derivative (HQ3) having a general structural formula as shown in Formula I-2-3:

[0143]

[0144] The specific preparation route is:

[0145]

[0146] The structural identification data is: 2-phenyl-5,7-dihydroxy-6-((6-(N-pyrimidin-2-yl-piperazine)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (HQ3): 1H NMR (400MHz, DMSO-d6) δ12.88 (s, 1H), 8.39 (d, J = 4.7Hz, 2H), 8.07 (d, J = 6.9Hz, 2H), 7.76-7.44 (m, 3H), 6.95 (s, 1H), 6.66 (t, J=4.7Hz, 1H), 6.58 (s, 1H), 6.10 (s, 1H), 3.94-3.72 (m, 4H), 3.73-3.58 (m, 4H), 2.23 (s, 3H); 13 C NMR (101MHz, DMSO-d6) δ184.74, 167.00, 166.01, 163.95, 161.13, 157.93, 153.95, 134.34, 131.87, 13 0.89, 129.48, 129.11, 126.35, 124.64, 110.33, 104.73, 94.67, 82.49, 43.29, 42.83, 25.78; IR(KBr)ν max (cm -1 ): 3429, 3067, 2855, 1663, 1585, 1450, 1190, 983, 798, 688, 575; ESI-MS displays a peak at m / z 526.0, calcd for [M+H] + :525.5445.

[0147] Example 12

[0148] The twelfth embodiment of the present invention provides a flavonoid derivative (HQ4) having a general structural formula as shown in Formula I-2-4:

[0149]

[0150] The specific preparation route is:

[0151]

[0152] The difference from Example 9 is that: fatty amine The baicalein derivative HQ4 was generated in 80% yield.

[0153] The structural identification data is: 2-phenyl-5,7-dihydroxy-6-((6-(N-methylpiperazine)-2-methylpyrimidin-4-yl))oxy)-4H-chromen-4-one (HQ4): 1H NMR (400MHz, CDCl3) δ13.02 (s, 1H), 7.98-7.80 (m, 2H), 7.66-7.44 (m, 3H), 6.66 (s, 1H), 6 .63(s, 1H), 6.00(s, 1H), 3.77-3.60(m, 4H), 2.67-2.44(m, 4H), 2.38(s, 3H), 2.35(s, 3H); 13 C NMR (101MHz, DMSO-d6) δ182.06, 169.56, 165.98, 163.97, 163.29, 158.13, 153.76, 152.61, 131.97, 1 30.80, 129.12, 126.42, 124.38, 104.81, 103.83, 94.45, 82.44, 54.15, 45.62, 43.50, 25.76; IR(KBr)ν max (cm -1 ): 3430, 3069, 2934, 1663, 1623, 1603, 1452, 1194, 990, 814, 772, 688; ESI-MS displays a peak at m / z 462.0, calcd for [M+H] + :461.4975.

[0154] Comparative Example 1

[0155] The first comparative example of the present invention is the anticancer drug cisplatin.

[0156] Comparative Example 2

[0157] The second comparative example of the present invention is a chrysin intermediate.

[0158] Comparative Example 3

[0159] The third comparative example of the present invention is a baicalein intermediate.

[0160] The flavonoid derivatives synthesized above were used to evaluate the inhibition rate and IC of the compounds in Examples 1 to 12 and Comparative Examples 1 to 3 on the growth of different human cancer cells using the MTS experimental research method. 50 value.

[0161] The MTS method was used to test the cytotoxicity of the compounds in Examples 1 to 12 and Comparative Examples 1 to 3 at a concentration of 40 μM against five human cancer cell lines: human lung cancer cells (A549), human breast cancer cells (MDA-MB-231), human liver cancer cells (HepG2), human colon cancer cells (HCT116), and human prostate cancer cells (PC-3). For products with good activity, the corresponding IC 50 value.

[0162] Principle of the MTS assay for cell viability: MTS, a new MTT analogue (3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazoliu m), is a yellow dye. Succinate dehydrogenase in the mitochondria of living cells metabolizes and reduces MTS to form a soluble formazan compound. The formazan content can be measured using a microplate reader at 490 nm. Generally, the amount of formazan produced is proportional to the number of viable cells, so the number of viable cells can be estimated based on the optical density (OD) value.

[0163] Activity test experimental method

[0164] 1) Cell inoculation: Prepare a single cell suspension in culture medium (DMEM or RMPI1640) containing 10% fetal bovine serum, and seed 3,000 to 5,000 cells per well in a 96-well plate with a volume of 100 μL per well. Cells should be inoculated and cultured 12 to 24 hours in advance.

[0165] 2) Add the test compound solution: dissolve the compound in DMSO or 0.5N dilute hydrochloric acid, initially screen the compound at a concentration of 40 μM, and add 200 μL of the final volume per well. Set up 3 replicate wells for each treatment.

[0166] 3) Color Development: After 48 hours of incubation at 37°C, discard the culture medium from the adherent cells and add 20 μL of MTS solution and 100 μL of culture medium to each well. Set up three blank replicate wells (a mixture of 20 μL of MTS solution and 100 μL of culture medium) and continue incubation for 2-4 hours to allow the reaction to proceed fully before measuring the absorbance.

[0167] 4) Colorimetry: Select 492 nm wavelength and read the absorbance of each well with a multifunctional microplate reader. Record the results. After data processing, plot a cell inhibition rate graph with the compound number as the horizontal axis and the cell inhibition rate as the vertical axis.

[0168] 5) Positive control compound: The initial screening concentration was 40 μM. A cell growth curve was drawn with the concentration as the horizontal axis and the cell viability as the vertical axis. The IC value of the compound was calculated using the two-point method.50 value.

[0169] Inhibitory rate and IC of the target compound against five cancer cells at a concentration of 40 μM 50 Values ​​(μM) are shown in Table 1 (Note: IC 50 Only compounds with good anticancer activity were determined).

[0170] Table 1:

[0171]

[0172] Note:(%) a is the inhibition rate of the target compound on five cancer cells at a concentration of 40 μM; IC 50 (μM) b The values ​​are the concentrations required to inhibit 50% of cancer cell apoptosis; the data from three repeated experiments are expressed as mean ± SD.

[0173] According to Examples 1 to 12 and Comparative Examples 1 to 3, the flavonoid derivatives have an inhibitory effect on human cancer cells.

[0174] Among them, the inhibitory activity of chrysin derivative BY6 on A549 cell line was higher than that of anticancer drug cisplatin, IC 50 The inhibitory activity against HCT116 and PC-3 cell lines was comparable to that of the anticancer drug cisplatin, with an IC 50 The values ​​were 19.47 ± 0.51 μM and 9.18 ± 0.25 μM, respectively.

[0175] Throughout this specification, reference to terms such as "one embodiment," "some embodiments," "examples," "specific examples," or "some examples" means that a specific feature, structure, material, or characteristic described in conjunction with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0176] The above-described embodiments merely illustrate several implementations of the present invention. While the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that a person skilled in the art would be able to make numerous modifications and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the present invention shall be determined by the appended claims.

Claims

1. A flavonoid derivative, characterized in that: The flavonoid derivatives include chrysin derivatives, and the general structural formula of the chrysin derivatives is shown in Formula I-1: in, 2. A method for preparing a flavonoid derivative, characterized in that: The preparation method is used to prepare the flavonoid derivative according to claim 1, and the preparation method comprises: reacting a flavonoid compound with 4,6-dichloro-2-methylpyrimidine to generate an intermediate, wherein the flavonoid compound includes chrysin; The intermediate is reacted with a corresponding fatty amine to obtain a flavonoid derivative.

3. The method for preparing the flavonoid derivative according to claim 2, characterized in that: The synthetic route of the flavonoids is as follows: Among them, fatty amine Dissolving chrysin and 4,6-dichloro-2-methylpyrimidine in an organic solvent DMF at a first predetermined molar ratio, and stirring the mixture under the action of an alkaline reagent K2CO3 to obtain a chrysin intermediate; The chrysin intermediate and the corresponding fatty amine 1 are dissolved in a solvent ethanol at a second preset molar ratio, and the temperature is raised to a preset temperature and reacted for a preset time under the action of an alkaline reagent DIPEA to obtain a chrysin derivative.

4. The method for preparing the flavonoid derivative according to claim 3, characterized in that: The first preset molar ratio is 1:(0.7-1.3), and the second preset molar ratio is 1:(0.9-1.5).

5. The method for preparing the flavonoid derivative according to claim 3, characterized in that: The preset temperature is 70° C.-90° C., and the preset time is 9 hours-12 hours.

6. The method for preparing the flavonoid derivative according to claim 3, characterized in that: The molar amount of the alkaline reagent K2CO3 is 10 mmol-15 mmol, the volume of the organic solvent DMF is 10 mL-50 mL, the molar amount of the alkaline reagent DIPEA is 1 mmol-1.5 mmol, and the volume of the ethanol is 1 mL-10 mL.

7. The method for preparing a flavonoid derivative according to claim 3, wherein: After the step of stirring the reaction under the action of the alkaline reagent K2CO3, the method further comprises: Deionized water was added and stirred for 5-10 minutes, the filtrate was removed by filtration, and the filter cake was subjected to column chromatography to obtain a chrysin intermediate.

8. The method for preparing a flavonoid derivative according to claim 3, wherein: After the step of heating to a preset temperature and reacting for a preset time under the action of the alkaline reagent DIPEA, the method further comprises: After the reaction is completed, the mixture is concentrated in vacuo, cooled to room temperature, and washed with ethyl acetate or petroleum ether and recrystallized to obtain a chrysin derivative.

9. Use of the flavonoid derivative according to claim 1 in the preparation of anticancer drugs.

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