Diphyllin urea derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicinal chemistry and pharmacology, and particularly relates to a diphyllin urea derivative as well as a preparation method and application thereof. The structural formula of the diphyllin urea derivative is shown in the specification, wherein R represents one of phenyl, 4-fluorophenyl, 4-chlorphenyl, 4-bromophenyl, 4-methylphenyl, 4-methoxyphenyl, 3-trifluoromethylphenyl, 2-chlorphenyl and 2-fluorophenyl. The structural formula of the diphyllin urea derivative is shown in the specification, and R represents one of phenyl, 4-fluorophenyl, 4-chlorphenyl, 4-bromophenyl, 4-methylphenyl, 4-methoxyphenyl, 3-trifluoromethylphenyl, 2-chlorphenyl and 2-fluorophenyl. The structure of the diphyllin urea derivative does not contain glycosidic bonds, double bonds and other structures which are easy to hydrolyze in vivo, the metabolic stability of the diphyllin urea derivative is superior to that of diphyllin glycoside compounds, and the diphyllin urea derivative has high tumor cell proliferation inhibition activity and can be applied to preparation of drugs for preventing and treating liver cancer and lung cancer.

Description

Technical Field

[0001] This invention belongs to the fields of medicinal chemistry and pharmacology, specifically relating to a urea derivative of senna leaf extract, its preparation method, and its application. Background Technology

[0002] Cancer is a major public health problem that seriously threatens human health, and the discovery and optimization of antitumor lead compounds from plants is one of the important research directions in this field. Cleistanthin, a natural lignan extracted from plants of the genus *Cleistanthin* in the Berberidaceae family, is a key pharmacodynamic skeleton for many bioactive derivatives. While this compound itself possesses antiviral activities, its direct antitumor effect is limited. Subsequent studies have found that cleistanthin glycoside derivatives (such as cleistanthin-A) exhibit excellent antitumor activity, showing promising development prospects (Chinese Journal of Chemistry, 2007, 25, 679-682; Chemical Biology and Drug Design, 2015, 86, 691-696). However, these glycoside derivatives not only have complex synthetic routes, but their glycosidic bonds are also easily hydrolyzed by enzymes in vivo, leading to poor metabolic stability.

[0003] To improve stability, researchers further designed and synthesized 4-C-linked kaempferol triazole derivatives. Unfortunately, while these new derivatives showed significantly improved metabolic stability, their antitumor activity was drastically reduced (Chemical Biology & Drug Design, 2024, 104, e14635). This result highlights the challenge of balancing activity and pharmacokinetic properties in lead compound optimization.

[0004] In medicinal chemistry, introducing specific, advantageous pharmacophores is a key strategy for overcoming such challenges. Besides common amide and sulfonamide groups, the urea structure is also of central importance. It can act as an excellent hydrogen bond donor and acceptor, forming strong and specific interactions with biological targets, thereby significantly improving the binding affinity and selectivity of compounds. Simultaneously, this structure also plays a positive role in regulating the physicochemical properties, solubility, and metabolic stability of molecules. Therefore, introducing the urea structure or its analogues into the optimization of the senna leaf extract structure can simultaneously overcome the limitations of its derivatives in terms of activity and stability.

[0005] Summary of the Invention

[0006] In view of this, the purpose of the present invention is to provide a urea derivative of kaempferol, its preparation method and application. Compared with natural kaempferol glycosides, the urea derivative of kaempferol does not contain glycosidic bonds, double bonds and other structures that are easily hydrolyzed in vivo. It has better metabolic stability than kaempferol glycoside derivatives and has stronger tumor cell proliferation inhibitory activity.

[0007] In a first aspect, the present invention provides a kaempferol urea derivative having the structure shown in the following formula:

[0008]

[0009] Wherein, R represents one of phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-methylphenyl, 4-methoxyphenyl, 3-trifluoromethylphenyl, 2-chlorophenyl and 2-fluorophenyl.

[0010] In some embodiments of the present invention, the kaempferol urea derivatives have a structure as shown in any one of formulas 4a-4e:

[0011]

[0012] in,

[0013] When R is phenyl, the safflower leaf urea derivative is a compound with the structure shown in formula 4a;

[0014] When R is p-fluorophenyl, the kaempferol urea derivative is a compound with the structure shown in formula 4b;

[0015] When R is p-methylphenyl, the safflower urea derivative is a compound with the structure shown in formula 4c;

[0016] When R is 3-trifluoromethylphenyl, the safflower urea derivative is a compound with the structure shown in formula 4d;

[0017] When R is 2-chlorophenyl, the kaempferol urea derivative is a compound with the structure shown in formula 4e.

[0018] A second aspect of the present invention provides a method for preparing kaempferol urea derivatives, comprising the following steps:

[0019] S1. The reaction of salifoliol with epibromopropane and potassium carbonate in N,N-dimethylformamide yields salifoliol epoxy ether 1.

[0020] S2. The reaction of kaempferol epoxy ether 1 with sodium azide and ammonium chloride in a mixed solution of N,N-dimethylformamide and water yields kaempferol ring-opening derivative 2.

[0021] S3. The open-ring derivative 2 of senna leaf extract undergoes a Staudinger reaction with triphenylphosphine to give the amino derivative 3 of senna leaf extract;

[0022] S4. The amino derivative 3 of senna leaf extract and benzyl isocyanate compound 5 undergo a substitution reaction in the presence of N,N-diisopropylethylamine to give urea derivative 4 of senna leaf extract;

[0023] The reaction formula for the above preparation method is as follows:

[0024] ,

[0025] Wherein, R represents one of phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-methylphenyl, 4-methoxyphenyl, 3-trifluoromethylphenyl, 2-chlorophenyl and 2-fluorophenyl.

[0026] In some embodiments of the present invention, the above-described preparation method includes the following steps:

[0027] (1) Dissolve senna leaf extract in N,N-dimethylformamide and stir at 25°C for 5 minutes. Slowly add epoxybromopropane and potassium carbonate to the mixed solution to obtain the first reaction solution. Quench the reaction with deionized water, extract with ethyl acetate, collect the organic phase and wash it with water, saturated brine, dry it with anhydrous MgSO4, concentrate it under reduced pressure, and then flash column chromatography to obtain a pale yellow solid, namely senna leaf extract epoxy ether 1;

[0028] (2) Dissolve the 1-hydroxy ether of senna leaf extract and ammonium chloride in a mixed solution of DMF:H2O, add ammonium chloride, heat to react, and obtain a second reaction solution. Add deionized water to the second reaction solution to quench the reaction, extract with ethyl acetate, collect the organic phase, wash with saturated brine, dry with anhydrous MgSO4, concentrate under reduced pressure, and obtain a white solid by flash column chromatography, namely the 2-cyclic compound of senna leaf extract.

[0029] (3) Dissolve the open-ring compound 2 of senna leaf extract in tetrahydrofuran solution, add triphenylphosphine and water, heat to react, and obtain a third reaction solution. After cooling the third reaction solution, concentrate it under reduced pressure, and then flash column chromatography to obtain a white solid, namely senna leaf extract amino derivative 3.

[0030] (4) Add the amino derivative 3 of senna leaf extract and the benzyl isocyanate compound 5 to the reaction tube. Under nitrogen protection, add N,N-diisopropylethylamine and react at room temperature to obtain the fourth reaction solution. Concentrate under reduced pressure and then flash column chromatography to obtain a white solid, namely the urea derivative 4 of senna leaf extract.

[0031] In some embodiments of the present invention, in step S1 of the above preparation method, the molar ratio of senna leaf extract, epichlorohydrin and potassium carbonate is 1:5:6; the reaction temperature is 40℃-70℃; and the reaction time is 1-3 hours.

[0032] In some embodiments of the present invention, in step S2 of the above preparation method, the molar ratio of senna leaf extract epoxy ether 1, sodium azide, and ammonium chloride is 1:3:2.

[0033] In some embodiments of the present invention, in step S2 of the above preparation method, the reaction temperature is 40℃-70℃ and the reaction time is 12 hours.

[0034] In some embodiments of the present invention, in step S3 of the above preparation method, the molar ratio of 2-ring-opening derivative of senna leaf extract and triphenylphosphine is 1:3.

[0035] In some embodiments of the present invention, in step S3 of the above preparation method, the reaction temperature is 40℃-80℃ and the reaction time is 3 hours.

[0036] In some embodiments of the present invention, in step S4 of the above preparation method, the molar ratio of kaempferol amino derivative 3, benzyl isocyanate compound, and N,N-diisopropylethylamine is 1:1.2:2.0.

[0037] In some embodiments of the present invention, in step S4 of the above preparation method, the reaction temperature is 20℃-50℃ and the reaction time is 12 hours.

[0038] In a third aspect, the present invention provides the use of the above-mentioned senna leaf extract urea derivatives in the preparation of a medicament for treating cancer, wherein the cancer is liver cancer or lung cancer.

[0039] Compared with existing technologies, this invention provides a urea derivative of kaempferol with a non-glycosidic structure. These compounds do not contain glycosidic bonds that are easily hydrolyzed in vivo, and their metabolic stability is superior to that of glycosidic compounds. Furthermore, in vitro tumor cell proliferation inhibition experiments have shown that these compounds possess strong tumor cell proliferation inhibitory activity, comparable to or superior to the positive control drug paclitaxel, and can be applied to the preparation of drugs for the prevention and treatment of liver and lung cancer. Attached Figure Description

[0040] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.

[0041] Figure 1 Nuclear magnetic resonance of 4a provided in Embodiment 3 of the present invention 1 H spectrum;

[0042] Figure 2 Nuclear magnetic resonance of 4a provided in Embodiment 3 of the present invention 13 C spectrum;

[0043] Figure 3 The nuclear magnetic resonance of 4b provided in Embodiment 4 of the present invention 1 H spectrum;

[0044] Figure 4 The nuclear magnetic resonance of 4b provided in Embodiment 4 of the present invention 13 C spectrum;

[0045] Figure 5 4c nuclear magnetic resonance provided in Embodiment 5 of the present invention 1 H spectrum;

[0046] Figure 6 4c nuclear magnetic resonance provided in Embodiment 5 of the present invention 13 C spectrum;

[0047] Figure 7 4d nuclear magnetic resonance provided in Embodiment 6 of the present invention 1 H spectrum;

[0048] Figure 8 4d nuclear magnetic resonance provided in Embodiment 6 of the present invention 13 C spectrum;

[0049] Figure 9 The 4e nuclear magnetic resonance provided in Embodiment 7 of the present invention 1 H spectrum;

[0050] Figure 10 The 4e nuclear magnetic resonance provided in Embodiment 7 of the present invention 13 C-spectrum. Detailed Implementation

[0051] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0052] Example 1

[0053] Kaempferol (1.0 mmol), epichlorohydrin (5.0 mmol), and potassium carbonate (6.0 mmol) were dissolved in DMF and stirred at 60 °C for 2 h. The reaction was monitored by thin-layer chromatography. After the reaction was complete, the aqueous phase was extracted with ethyl acetate, and the organic phase was extracted with saturated sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate = 2:1) to give a yellow solid, kaempferol epoxy ether 2 (0.85 mmol, 85%). f = 0.5 (petroleum ether: ethyl acetate = 1:1). 1 H NMR (400 MHz, CDCl3) δ 7.63 (s, 1H, ArH), 7.07 (s, 1H, ArH), 6.96 (d, J= 7.8 Hz, 1H, ArH), 6.84-6.77 (m, 2H, ArH), 6.11-6.03 (m, 2H, OCH2O), 5.47(s, 2H, OCH2), 4.56 (dd, J = 11.2, 2.1 Hz, 1H, OCH2), 4.10 (s, 3H, OCH3), 4.07-4.00 (m, 1H, OCH2), 3.81 (s, 3H, OCH3), 3.46 (ddt, J = 6.6, 4.5, 2.4 Hz,1H, OCH2), 2.98 (t, J = 4.5 Hz, 1H, CH), 2.83 (dd, J = 4.9, 2.6 Hz, 1H, OCH2). 13 C NMR (101 MHz, CDCl3) δ 169.6, 151.8, 150.3, 147.5, 146.4, 135.4, 130.7,128.2, 127.4, 126.7, 123.6, 119.1, 110.7, 108.2, 106.2, 101.2, 100.4, 73.7,66.3, 56.2, 55.8, 50.4, 44.4.

[0054] Example 2

[0055] Kaempferol (1.0 mmol), sodium azide (3.0 mmol), and ammonium chloride (2.0 mmol) were dissolved in a mixed solution of DMF:H₂O = 4:1. The mixture was stirred at 70 °C for 12 h, and the reaction was monitored by thin-layer chromatography. After the reaction was complete, the aqueous phase was extracted with ethyl acetate, and the organic phase was extracted with saturated sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by column chromatography (petroleum ether:ethyl acetate = 2:1) to give a white solid kaempferol ring-opening derivative 2 (0.85 mmol, 87%). f = 0.5 (petroleum ether: ethyl acetate = 1:1); 1 H NMR (400 MHz, Chloroform-d) δ 7.58 (s, 1H, ArH), 7.06 (s, 1H, ArH), 6.93 (d, J = 7.8 Hz, 1H, ArH), 6.81 – 6.71 (m, 2H, ArH), 6.05 (dd, J = 19.6, 1.4 Hz, 2H, OCH2O), 5.44 (s, 2H, COOCH2), 4.28 (p, J = 5.2Hz, 1H, HOCH), 4.21 (d, J = 4.6 Hz, 2H, OCH2), 4.06 (s, 3H, OCH3), 3.80 (s,3H, OCH3), 3.64 (qd, J = 12.6, 5.3 Hz, 2H, N3CH2). 13 C NMR (100 MHz, CDCl3) δ169.8, 151.9, 150.4, 147.5, 146.3, 135.5, 130.8, 128.2, 127.1, 126.6, 123.6,119.1, 110.7, 108.2, 106.3, 101.3, 100.3, 73.7, 69.7, 66.4, 56.2, 55.9, 53.3.HRMS (ESI): m / z calcd for C 24 H 21 N3O8: 480.1407; found: 480.1411 [M + H] +

[0056] Example 3

[0057] Compound 2 (1.0 mmol) was dissolved in tetrahydrofuran solution, and triphenylphosphine (3.0 mmol) and H₂O (10.0 mmol) were added. The reaction temperature was raised to 75°C and stirred for 3 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the aqueous phase was extracted with ethyl acetate, and the organic phase was extracted with saturated sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate = 1:1) to give a yellow solid, kaempferol amino derivative 3 (0.85 mmol, 85%). f = 0.5 (petroleum ether: ethyl acetate = 1:1); Under a nitrogen atmosphere, at 0°C, 1.0 mmol of kaempferol amino derivative 3 was dissolved in 5 mL of anhydrous dichloromethane solution, and N,N-diisopropylethylamine (2.0 mmol) was added. After stirring for 5 minutes, an anhydrous dichloromethane solution of benzyl isocyanate (1.2 mmol) was slowly added dropwise. The mixture was stirred at this temperature for 30 minutes, and then the reaction temperature was raised to room temperature and stirred for 12 hours. After the reaction was completed as monitored by thin-layer chromatography, a large amount of water was added to the reaction solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated by column chromatography (dichloromethane: ethyl acetate = 2:1) to give a white solid 4a (0.77 mmol, 77%). R f =0.5 (dichloromethane:methanol = 12:1). 1 H NMR (400MHz, CDCl3) δ 7.60 (s, 1H, ArH), 7.25 (s, 4H, ArH), 7.10 – 7.05 (m, 2H, ArH), 6.93 (dd, J = 7.8, 2.6 Hz, 1H, ArH), 6.81 – 6.75 (m, 2H, ArH), 6.04 (dd, J =21.6, 2.9 Hz, 2H, H-7'), 5.39 (d, J = 3.3 Hz, 2H, H-3a), 4.21 (dq, J = 7.6,4.5, 3.5 Hz, 1H, H-2''), 4.13 (d, J = 6.9 Hz, 2H, H-5''), 4.13 – 4.05 (m, 2H,H-1''), 4.02 (s, 3H, -OCH3), 3.80 (s, 3H, -OCH3), 3.70 – 3.66 (m, 1H, H-3''), 3.56 (dt, J = 13.7, 5.6 Hz, 1H, H-3''). 13C NMR (100MHz, CDCl3) δ 169.8,157.8, 156.5, 151.8, 150.3, 147.5, 146.5, 137.8, 136.6, 130.7, 129.2, 128.7,127.5, 127.2, HRMS (ESI): m / z calcd forC 32 H 30 N2O9Na: 609.1843; found: 609.1849 [M+Na] +

[0058] Nuclear magnetic resonance of white solid 4a 1 H spectrum as shown Figure 1 As shown; nuclear magnetic resonance 13 C spectrum as shown Figure 2 As shown.

[0059] Example 4

[0060] Compound 2 (1.0 mmol) was dissolved in tetrahydrofuran solution, and triphenylphosphine (3.0 mmol) and H₂O (10.0 mmol) were added. The reaction temperature was raised to 75°C and stirred for 3 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the aqueous phase was extracted with ethyl acetate, and the organic phase was extracted with saturated sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate = 1:1) to give a yellow solid, kaempferol amino derivative 3 (0.85 mmol, 85%). f = 0.5 (petroleum ether: ethyl acetate = 1:1); Under a nitrogen atmosphere, at 0°C, 1.0 mmol of kaempferol amino derivative 3 was dissolved in 5 mL of anhydrous dichloromethane solution, and N,N-diisopropylethylamine (2.0 mmol) was added. After stirring for 5 minutes, an anhydrous dichloromethane solution of benzyl p-fluoroisocyanate (1.2 mmol) was slowly added dropwise. The mixture was stirred at this temperature for 30 minutes, and then the reaction temperature was raised to room temperature and stirred for 12 hours. After the reaction was completed as monitored by thin-layer chromatography, a large amount of water was added to the reaction solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated by column chromatography (dichloromethane: ethyl acetate = 2:1) to give a white solid 4b (0.73 mmol, 73%). R f =0.4 (dichloromethane:methanol = 12:1). 1H NMR(400 MHz, CDCl3) δ 7.59 (s, 1H, ArH), 7.13 – 7.08 (m, 2H, ArH), 7.04 (d, J =2.0 Hz, 1H, ArH), 6.96 – 6.89 (m, 3H, ArH), 6.80 – 6.72 (m, 2H, ArH), 6.03(dt, J = 22.4, 1.6 Hz, 2H, H-7'), 5.39 (s, 2H, H-3a), 4.38 – 4.32 (m, 2H, H-5''), 4.11 (d, J = 7.1 Hz, 2H, H-1''), 4.09 (s, 1H, H-2''), 4.02 (s, 3H, -OCH3), 3.79 (s, 3H, -OCH3), 3.70 – 3.63 (m, 1H, H-3''), 3.58 – 3.49 (m, 1H, H-3''). 13 C NMR (100 MHz, CDCl3) δ 171.2, 163.3, 157.6, 156.3, 151.8, 147.5,135.2, 130.7, 129.0, 129.0, 128.0, 127.9, 126.5, 123.6, 119.0, 116.2, 115.4,110.7, 108.2, 101.3, 74.4, 70.2, 66.6, 60.4, 56.1, 55.8, 46.0, 44.0. HRMS(ESI): m / z calcd for C 32 H 29 N2O9NaF: 627.1747; found: 627.1755 [M+Na] +

[0061] Nuclear magnetic resonance of white solid 4b 1 H spectrum as shown Figure 3 As shown; nuclear magnetic resonance 13 C spectrum as shown Figure 4 As shown.

[0062] Example 5

[0063] Compound 2 (1.0 mmol) was dissolved in tetrahydrofuran solution, and triphenylphosphine (3.0 mmol) and H₂O (10.0 mmol) were added. The reaction temperature was raised to 75°C and stirred for 3 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the aqueous phase was extracted with ethyl acetate, and the organic phase was extracted with saturated sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate = 1:1) to give a yellow solid, kaempferol amino derivative 3 (0.85 mmol, 85%). f = 0.5 (petroleum ether: ethyl acetate = 1:1); Under a nitrogen atmosphere, at 0°C, 1.0 mmol of kaempferol amino derivative 3 was dissolved in 5 mL of anhydrous dichloromethane solution, and N,N-diisopropylethylamine (2.0 mmol) was added. After stirring for 5 minutes, an anhydrous dichloromethane solution of benzyl p-methylisocyanate (1.2 mmol) was slowly added dropwise. The mixture was stirred at this temperature for 30 minutes, and then the reaction temperature was raised to room temperature and stirred for 12 hours. After the reaction was completed as monitored by thin-layer chromatography, a large amount of water was added to the reaction solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated by column chromatography (dichloromethane: ethyl acetate = 2:1) to give a white solid 4c (0.65 mmol, 65%). R f =0.2 (dichloromethane:methanol = 12:1). 1 H NMR(400 MHz, CDCl3) δ 7.61 (s, 1H, ArH), 7.05 (dd, J = 4.1, 1.8 Hz, 2H, ArH), 6.99 – 6.94 (m, 3H, ArH), 6.93 (d, J = 2.2 Hz, 1H, ArH), 6.78 (ddd, J = 10.5,5.3, 2.4 Hz, 2H, ArH), 6.04 (ddd, J = 21.5, 3.1, 1.6 Hz, 2H, H-7'), 5.41 –5.38 (d, 2H, H-3a), 4.32 (d, J = 5.3 Hz, 2H, H-5''), 4.26 – 4.20 (m, 1H, H-2''), 4.10 (d, J = 7.1 Hz, 2H, H-1''), 4.02 (s, 3H, -OCH3), 3.80 (s, 3H, -OCH3), 3.73 – 3.71 (m, 1H, H-3''), 3.60 – 3.52 (m, 1H, H-3''), 2.31 (d, J =1.2 Hz, 3H, -CH3). 13C NMR (100 MHz, CDCl3) δ 171.2, 156.6, 151.7, 150.3,146.5, 137.1, 133.5, 129.9, 129.3, 128.4, 127.2, 126.2, 119.1, 110.7, 108.2,101.3, 74.4, 70.4, 66.6, 60.4, 56.2, 55.8, 46.5, 44.5, 21.1. HRMS (ESI): m / zcalcd for C 33 H 32 N2O9Na: 623.1994; found: 623.2006 [M+Na] +

[0064] Nuclear magnetic resonance of white solid 4C 1 H spectrum as shown Figure 5 As shown; nuclear magnetic resonance 13 C spectrum as shown Figure 6 As shown.

[0065] Example 6

[0066] Compound 2 (1.0 mmol) was dissolved in tetrahydrofuran solution, and triphenylphosphine (3.0 mmol) and H₂O (10.0 mmol) were added. The reaction temperature was raised to 75°C and stirred for 3 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the aqueous phase was extracted with ethyl acetate, and the organic phase was extracted with saturated sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate = 1:1) to give a yellow solid, kaempferol amino derivative 3 (0.85 mmol, 85%). f = 0.5 (petroleum ether: ethyl acetate = 1:1); Under a nitrogen atmosphere, at 0°C, 1.0 mmol of kaempferol amino derivative 3 was dissolved in 5 mL of anhydrous dichloromethane solution, and N,N-diisopropylethylamine (2.0 mmol) was added. After stirring for 5 minutes, an anhydrous dichloromethane solution of 1.2 mmol of 3-trifluoromethyl isocyanate was slowly added dropwise. The mixture was stirred at this temperature for 30 minutes, and then the reaction temperature was raised to room temperature and stirred for 12 hours. After the reaction was completed as monitored by thin-layer chromatography, a large amount of water was added to the reaction solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated by column chromatography (dichloromethane: ethyl acetate = 2:1) to give a white solid 4d (0.82 mmol, 82%). f =0.4 (dichloromethane:methanol = 12:1). 1HNMR (400 MHz, CDCl3) δ 7.58 (s, 1H, ArH), 7.51 – 7.43 (m, 3H, ArH), 7.33 (d,J = 7.9 Hz, 1H, ArH), 7.01 (s, 1H, ArH), 6.92 – 6.85 (m, 1H, ArH), 6.75 – 6.67 (m, 2H, ArH), 6.00 (dt, J = 23.9, 1.3 Hz, 2H, H-7'), 5.37 (s, 2H, H-3a), 4.48 – 4.43 (m, 2H, H-5''), 4.20 (dd, J = 7.6, 3.7 Hz, 1H, H-2''), 4.11 –4.04 (m, 2H, H-1''), 4.01 (s, 3H, -OCH3), 3.78 (s, 3H, -OCH3), 3.70 – 3.64 (m,1H, H-3''), 3.50 (dt, J = 13.9, 6.0 Hz, 1H, H-3''). 13 C NMR (100MHz, CDCl3) δ171.3, 157.4, 156.2, 150.3, 147.5, 146.4, 139.2, 137.8, 130.6, 130.6, 129.6,129.5, 129.1, 128.3, HRMS (ESI): m / z calcdfor C 33 H 30 N2O9F3: 655.1899; found: 655.1903 [M+Na] +

[0067] 4D nuclear magnetic resonance of white solid 1 H spectrum as shown Figure 7 As shown; nuclear magnetic resonance 13 C spectrum as shown Figure 8 As shown.

[0068] Example 7

[0069] Compound 2 (1.0 mmol) was dissolved in tetrahydrofuran solution, and triphenylphosphine (3.0 mmol) and H₂O (10.0 mmol) were added. The reaction temperature was raised to 75°C and stirred for 3 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the aqueous phase was extracted with ethyl acetate, and the organic phase was extracted with saturated sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate = 1:1) to give a yellow solid, kaempferol amino derivative 3 (0.85 mmol, 85%). f = 0.5 (petroleum ether: ethyl acetate = 1:1); Under a nitrogen atmosphere, at 0°C, 1.0 mmol of kaempferol amino derivative 3 was dissolved in 5 mL of anhydrous dichloromethane, and N,N-diisopropylethylamine (2.0 mmol) was added. After stirring for 5 minutes, an anhydrous dichloromethane solution of benzyl 2-chloroisocyanate (1.2 mmol) was slowly added dropwise. The mixture was stirred at this temperature for 30 minutes, and then the reaction temperature was raised to room temperature and stirred for 12 hours. After the reaction was completed as monitored by thin-layer chromatography, a large amount of water was added to the reaction solution and extracted with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated by column chromatography (dichloromethane: ethyl acetate = 2:1) to give a white solid 4e (0.79 mmol, 79%). f =0.2 (dichloromethane:methanol = 10:1). 1H NMR(400 MHz, CDCl3 ) δ 7.59 (s, 1H, ArH), 7.29 (d, J = 7.9 Hz, 1H, ArH), 7.23(s, 1H, ArH), 7.16 (d, J = 2.8 Hz, 2H, ArH), 7.06 (d, J = 2.8 Hz, 1H, ArH), 6.94 (dd, J = 7.6, 3.2 Hz, 1H, ArH), 6.82 – 6.76 (m, 2H, ArH), 6.06 (dq, J =18.7, 1.5 Hz, 2H, H-7'), 5.40 (d, J = 3.3 Hz, 2H, H-3a), 4.47 (dd, J = 5.7,1.4 Hz, 2H, H-5''), 4.24 (q, J = 5.2 Hz, 1H, H-2''), 4.13 (d, J = 7.4 Hz, 2H,H-1''), 4.02 (d, J = 2.3 Hz, 3H, -OCH3), 3.80 (d, J = 1.9 Hz, 3H, -OCH3), 3.71(ddd, J = 9.5, 6.6, 3.4 Hz, 1H, H-3''), 3.64 – 3.55 (m, 1H, H-3''). 13 C NMR(100MHz, CDCl3) δ 171.2, 157.5, 156.1, 150.3, 147.5, 146.4, 133.5, 132.2,130.7, 129.9, 129.7, 129.6, 129.0, 128.3, HRMS(ESI): m / z calcd for C 32 H 29 N2O9NaCl: 643.1450; found: 643.1459 [M+Na] +

[0070] Nuclear magnetic resonance of white solid 4e 1 H spectrum as shown Figure 9 As shown; nuclear magnetic resonance 13 C spectrum as shown Figure 10 As shown.

[0071] To better understand the essence of this invention, the following pharmacological experimental results demonstrating the inhibitory effect of the senna leaf extract urea derivatives provided by this invention on the growth of three tumor cell lines illustrate its novel application in the field of antitumor drug research. The pharmacological examples provide partial activity data for representative compounds. It must be noted that the pharmacological examples of this invention are for illustrative purposes only and not for limiting the invention. Simple modifications made to this invention based on its essence are all within the scope of protection claimed by this invention.

[0072] Drug Experiment Example 1

[0073] Cytotoxic activity tests of compounds 4a-4e and paclitaxel against human hepatocellular carcinoma cells (HepG2), human lung cancer cells (PC-9), and human lung cancer cells (A549)

[0074] Human hepatocellular carcinoma cells (HepG2), human lung cancer cells (PC-9), and human lung cancer cells (A549) were cultured in RPMI-1640 complete medium containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 U / mL streptomycin at 37°C in a humidified air incubator containing 5% CO2.

[0075] Cells in the logarithmic growth phase were seeded at a density of 5 × 10³ cells per well in 96-well plates and cultured for 24 hours to allow for full cell adhesion. The test compound was dissolved in DMSO to prepare 1 × 10³ solution. -2 The stock solution was diluted with complete culture medium to the corresponding concentrations to obtain test compound solutions of different concentrations. After removing the original culture medium, culture medium containing different concentrations of compound 3a was added, with four parallel wells for each concentration, and the mixture was incubated for another 68 hours. After incubation, tetramethylazobium salt (MTT) solution was added to each well, and the mixture was incubated for another 4 hours. The culture medium was then discarded, and 150 µL of dimethyl sulfoxide was added to each well, followed by shaking for 10 min. Finally, the absorbance (A) value at 570 nm was measured using a microplate reader, and the half-maximal inhibitory concentration (IC50) was calculated, as shown in Table 1.

[0076] Table 1. Results of cytotoxic activity tests of compounds 4a-4e and paclitaxel.

[0077]

[0078] As shown in Table 1, the kaempferol urea derivatives provided by this invention possess significant biological activity. In vitro cytotoxic activity tests on three types of tumor cells—hepatocellular carcinoma (HepG2), human lung cancer cells (A549), and human lung cancer cells (PC-9)—demonstrate that these kaempferol urea derivatives inhibit tumor cell growth and have the potential to be developed into novel anti-tumor drugs. From the above pharmacological examples, we can see that these compounds exhibit strong cytotoxic activity against these three types of tumor cells. The cytotoxic activity of most compounds is close to that of the positive control paclitaxel, indicating their potential for development into anti-tumor drugs.

[0079] While some embodiments of the present general inventive concept have been shown and described, those skilled in the art will understand that changes may be made to these embodiments without departing from the principles and spirit of the present general inventive concept, the scope of which is defined by the claims and their equivalents.

Claims

1. A urea derivative of lycopodium, characterized in that, The structural formula of the urea derivative of kaempferol is shown below: ， Wherein, R represents one of phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-methylphenyl, 4-methoxyphenyl, 3-trifluoromethylphenyl, 2-chlorophenyl and 2-fluorophenyl.

2. The urea derivative of lycopodium according to claim 1, characterized in that, The kaempferol urea derivatives have structures as shown in any one of formulas 4a-4e:

3. A method for preparing the urea derivative of senna leaf extract as described in claim 1, characterized in that, Includes the following steps: S1. The reaction of salifoliol with epibromopropane and potassium carbonate in N,N-dimethylformamide yields salifoliol epoxy ether 1. S2. The reaction of kaempferol epoxy ether 1 with sodium azide and ammonium chloride in a mixed solution of N,N-dimethylformamide and water yields kaempferol ring-opening derivative 2. S3. The open-ring derivative 2 of senna leaf extract undergoes a Staudinger reaction with triphenylphosphine to give the amino derivative 3 of senna leaf extract; S4. The amino derivative 3 of senna leaf extract and benzyl isocyanate compound 5 undergo a substitution reaction in the presence of N,N-diisopropylethylamine to give urea derivative 4 of senna leaf extract; The reaction formula for the above preparation method is as follows: ， Wherein, R represents one of phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-methylphenyl, 4-methoxyphenyl, 3-trifluoromethylphenyl, 2-chlorophenyl and 2-fluorophenyl.

4. The preparation method according to claim 3, characterized in that, In step S1, the molar ratio of senna leaf extract, epichlorohydrin and potassium carbonate is 1:5:6; the reaction temperature is 40℃-70℃ and the reaction time is 1-3 hours.

5. The preparation method according to claim 3, characterized in that, In step S2, the molar ratio of salifolin epoxy ether 1, sodium azide, and ammonium chloride is 1:3:2; the reaction temperature is 40℃-70℃, and the reaction time is 12 hours.

6. The preparation method according to claim 3, characterized in that, In step S3, the molar ratio of 2-caryophyllein ring-opening derivative and triphenylphosphine is 1:

3.

7. The preparation method according to claim 3, characterized in that, In step S3, the reaction temperature is 40℃-80℃ and the reaction time is 3 hours.

8. The preparation method according to claim 3, characterized in that, In step S4, the molar ratio of kaempferol amino derivative 3, benzyl isocyanate compound 5, and N,N-diisopropylethylamine is 1:1.2:

2.

9. The preparation method according to claim 3, characterized in that, In step S4, the reaction temperature is 20℃-50℃ and the reaction time is 12 hours.

10. The use of the senna-based urea derivatives as described in claim 1 in the preparation of a medicament for treating cancer, wherein the cancer is liver cancer or lung cancer.

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