Beta-elemonic acid derivative as well as preparation method and application thereof

Abstract

The invention discloses a beta-elemonic acid derivative and a preparation method and application thereof, and belongs to the technical field of biological medicine, the structural formula of the beta-elemonic acid derivative is shown in the specification, the R1 group is one of 4-H, 4-Cl, 4-OCH3 and 3, 4, 5-(OCH3) 3; and R2 is one of 4-H, 4-F, 4-Cl and 4-OCH3. Tetracyclic triterpenoid beta-elemonic acid is used as a lead compound, a 1, 2, 3-triazole structure with anticancer activity is introduced, and the generated beta-elemonic acid derivative has low toxicity to human normal hepatocytes, high selectivity to tumor cells and remarkable anti-proliferation effect.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a β-elemene acid derivative, its preparation method, and its application. Background Technology

[0002] Cancer is one of the most destructive and disturbing diseases, affecting the lives of millions worldwide. Current clinical drugs for cancer treatment have limitations and side effects, including narrow therapeutic windows and low selectivity. Therefore, developing novel anticancer drugs with low toxicity to normal cells, enhanced antitumor activity, and relatively large therapeutic windows is an important direction in current antitumor research. In recent years, an increasing number of studies have demonstrated that triterpenoids possess anticancer activity against various human cancer cell lines and xenograft tumors. β-elemeneic acid, a tetracyclic triterpenoid isolated from frankincense, possesses anti-inflammatory, antitumor, and pro-apoptotic effects; however, its poor water solubility, short half-life, low bioavailability, and relatively low antitumor activity limit its application.

[0003] Molecular hybridization (MH) is considered a strategy in drug design and development that involves mixing pharmacophores from different bioactive molecules to produce a hybrid compound with better affinity and efficacy compared to the parent drug. This approach yields skeletons with improved selectivity, dual modes of action, and lower side effects. Structural modification of active natural products is an effective method for discovering potential active molecules, lead compounds, and new drugs. Summary of the Invention

[0004] The purpose of this invention is to provide a β-elemene acid derivative, its preparation method, and its application. Using tetracyclic triterpenoid β-elemene acid as a lead compound, a 1,2,3-triazole structure with anticancer activity is introduced. The resulting β-elemene acid derivative has low toxicity to normal human liver cells, high selectivity for tumor cells, and significant anti-proliferation effect.

[0005] To achieve the above objectives, the present invention provides a β-elemene acid derivative with the following structural formula:

[0006]

[0007] The structural formula of the R-based base is: One of the following: R1 group is one of 4-H, 4-Cl, 4-OCH3 and 3,4,5-(OCH3)3; R2 group is one of 4-H, 4-F, 4-Cl and 4-OCH3.

[0008] Preferably, when the R1 group is 4-H, the structural formula of the β-elemene acid derivative is:

[0009]

[0010] When the R1 group is 4-Cl, the structural formula of the β-elemene acid derivative is:

[0011]

[0012] When the R1 group is 4-OCH3, the structural formula of the β-elemene acid derivative is:

[0013]

[0014] When the R1 group is 3,4,5-(OCH3)3, the structural formula of the β-elemene acid derivative is:

[0015]

[0016] Preferably, when the R2 group is 4-H, the structural formula of the β-elemene acid derivative is:

[0017]

[0018] When the R2 group is 4-F, the structural formula of the β-elemene acid derivative is:

[0019]

[0020] When the R2 group is 4-Cl, the structural formula of the β-elemene acid derivative is:

[0021]

[0022] When the R2 group is 4-OCH3, the structural formula of the β-elemene acid derivative is:

[0023]

[0024] This invention provides a method for preparing β-elemene acid derivatives, comprising the following steps:

[0025] S1. Add β-elemene acid and alkyl bromide / chlorine derivative to anhydrous acetonitrile, then add anhydrous potassium carbonate, heat and stir to obtain a mixed solution;

[0026] S2. The mixed solution was extracted using dichloromethane as the extractant, and the organic phase was obtained after washing and drying.

[0027] S3. The organic phase was purified by column chromatography to obtain β-elemene acid derivative.

[0028] Preferably, the alkyl bromide / chlorine derivative is one of 4-(chloromethyl)-1-phenyl-1H-1,2,3-triazole, 4-(chloromethyl)-1-(4-chlorophenyl)-1H-1,2,3-triazole, 4-(chloromethyl)-1-(4-methoxyphenyl)-1H-1,2,3-triazole, 4-(chloromethyl)-1-(3,4,5-trimethoxyphenyl)-1H-1,2,3-triazole, 4-(bromomethyl)-1-benzyl-1H-1,2,3-triazole, 4-(bromomethyl)-1-(4-fluorobenzyl)-1H-1,2,3-triazole, 4-(bromomethyl)-1-(4-chlorobenzyl)-1H-1,2,3-triazole, and 4-(bromomethyl)-1-(4-methoxybenzyl)-1H-1,2,3-triazole.

[0029] Preferably, the molar ratio of β-elemene, alkyl bromide / chloride derivative and anhydrous potassium carbonate in S1 is 1:(1.5-2):(2-3).

[0030] Preferably, in S1, the heating temperature is 40℃-60℃, the heating time is 12h-24h, and the stirring time is 12h-24h.

[0031] Preferably, in S3, the eluent used for column chromatography purification is a mixed solution of petroleum ether and ethyl acetate; the volume ratio of the petroleum ether to the ethyl acetate is (5-20):1.

[0032] The present invention also provides an application of a β-elemene acid derivative, wherein the β-elemene acid derivative is used in the preparation of an antitumor drug.

[0033] The present invention also provides a β-elemene acid derivative composition comprising the aforementioned β-elemene acid derivative, and a pharmaceutically acceptable carrier or excipient.

[0034] Therefore, the β-elemene acid derivative, its preparation method, and its application provided by this invention have the following advantages compared with traditional technologies:

[0035] (1) β-elemene derivatives: Using tetracyclic triterpenoid β-elemene as a lead compound, the 1,2,3-triazole structure with anticancer activity was introduced. The resulting β-elemene derivatives showed low toxicity to normal human hepatocytes, high selectivity for tumor cells, and significant anti-proliferation effects. Among them, the β-elemene derivative with R1 group 4-H, namely β-elemene derivative J1, had a selectivity index of up to 51.95 for SiHa cells and 45.34 for HCT-116 cells, both higher than that of β-elemene.

[0036] (2) The IC50 of β-elemene derivative J1 on SiHa, HepG2, HCT-116 and A549 cells 50 The concentrations of 0.96 μM, 2.51 μM, 1.10 μM, and 3.18 μM, respectively, showed antiproliferative efficacy approximately 56.71 times, 24.5 times, 42.64 times, and 15.7 times higher than that of β-elemeneic acid, indicating that these compounds have the potential to be used as antitumor drugs. Furthermore, they can be combined with pharmaceutically acceptable pharmaceutical carriers or excipients to prepare β-elemeneic acid derivative compositions.

[0037] The technical solution of the present invention will be further described in detail below through embodiments. Detailed Implementation

[0038] The technical solution of the present invention will be further described below through embodiments.

[0039] Unless otherwise defined, the technical or scientific terms used in this invention shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0040] The structural formula of a β-elemene acid derivative provided by this invention is as follows:

[0041]

[0042] The structural formula of the R-based base is: One of the following: R1 group is one of 4-H, 4-Cl, 4-OCH3 and 3,4,5-(OCH3)3; R2 group is one of 4-H, 4-F, 4-Cl and 4-OCH3.

[0043] The specific implementation examples are as follows:

[0044] Example 1

[0045] A β-elemene acid derivative, specifically when the R1 group is 4-H, has the following structural formula:

[0046]

[0047] The preparation method of β-elemene acid derivative J1 includes the following steps:

[0048] S1. Add 1 mmol of β-elemeneic acid and 1.5 mmol of 4-(chloromethyl)-1-phenyl-1H-1,2,3-triazole to 5 mL of anhydrous acetonitrile, then add 2 mmol of anhydrous potassium carbonate (K2CO3), heat to 40 °C and stir for 24 h to obtain a mixed solution.

[0049] S2. Perform TCL detection on the mixed solution. After β-elemene acid has completely disappeared, pour the mixed solution into 10 mL of water, and then use 8 mL of dichloromethane (DCM) as the extractant to extract the mixed solution. Extract three times, and then wash with saturated sodium chloride and dry with anhydrous sodium sulfate to obtain the organic phase.

[0050] S3. The organic phase was purified by column chromatography by gradient elution with a mixture of petroleum ether and ethyl acetate in volume ratios of 20:1, 10:1 and 5:1 to obtain β-elemene acid derivative J1 in 55% yield.

[0051] The nuclear magnetic resonance (NMR) analysis of β-elemene acid derivative J1 yielded the following spectral data:

[0052] 1H NMR spectrum: 1 H NMR (300MHz, CDCl3) δ8.06 (s, 1H), 7.77–7.69 (m, 2H), 7.54 (t, J = 7.4Hz, 2H), 7.50–7.43 (m, 1H), 5.38 (d,J=12.6Hz,1H),5.26(d,J=12.7Hz,1H),5.03(t,J=6.4Hz,1H),2.58–2.39(m,2H),2.39–2.29(m,1 H),2.11-2.09(m,2H),2.01–1.90(m,2H),1.87–1.67(d,J=7.0Hz,5H),1.64(s,3H),1.62(s,2H),1.5 6(s,6H),1.47(s,3H),1.30(s,3H),1.08(s,3H),1.04(s,3H),0.98(s,3H),0.86(s,3H),0.75(s,3H).

[0053] Carbon NMR spectrum: 13 C NMR (75MHz, CDCl3) δ217.95,176.27,143.80,135.35,134.78,134.32,132.6 3,132.16,130.00(2C),123.47,122.13,121.62(2C),57.01,51.42,49.52,47 .75,47.25,47.04,43.86,37.09,35.48,34.50,32.61,29.43,28.77,27.37,26.85,26.66,25.93,25.65,24.21,21.19,21.10,20.17,19.75,17.53,15.99.

[0054] Electrospray ionization source-high resolution mass spectrometry (m / z): The calculated value of molecular protonated ion is 612.4160, and the measured value is 612.4165.

[0055] Example 2

[0056] The structural formula of a β-elemene acid derivative, specifically when the R1 group is 4-Cl, is as follows:

[0057]

[0058] The preparation method of β-elemene acid derivative J2 includes the following steps:

[0059] S1. Add 1 mmol of β-elemeneic acid and 1.5 mmol of 4-(chloromethyl)-1-(4-chlorophenyl)-1H-1,2,3-triazole to 5 mL of anhydrous acetonitrile, then add 2 mmol of anhydrous potassium carbonate (K2CO3), heat to 60 °C and stir for 12 h to obtain a mixed solution.

[0060] S2. Perform TCL detection on the mixed solution. After β-elemene acid has completely disappeared, pour the mixed solution into 10 mL of water, and then use 8 mL of dichloromethane (DCM) as the extractant to extract the mixed solution. Extract three times, and then wash with saturated sodium chloride and dry with anhydrous sodium sulfate to obtain the organic phase.

[0061] S3. The organic phase was purified by column chromatography by gradient elution with petroleum ether and ethyl acetate in volume ratios of 20:1, 10:1 and 5:1 to obtain β-elemene acid derivative J2 in 58% yield.

[0062] The nuclear magnetic resonance (NMR) analysis of β-elemene acid derivative J2 yielded the following spectral data:

[0063] 1H NMR spectrum: 1H NMR (300MHz, CDCl3) δ8.06(s,1H),7.69(d,J=8.8Hz,2H),7.52(d,J=8.8Hz,2H),5.32(dd,J=2 7.5,12.7Hz,2H),5.04(s,1H),2.63–2.51(m,1H),2.50–2.42(m,1H),2.36(td,J=14.4,7.5Hz ,1H),2.10(dd,J=18.1,10.3Hz,2H),2.03–1.77(m,7H),1.65(s,3H),1.63–1.59(m,6H),1.47 (s,3H),1.39–1.21(m,5H),1.10(s,3H),1.06(s,3H),1.01(s,3H),0.88(s,3H),0.77(s,3H).

[0064] Carbon NMR spectrum: 13 C NMR (75MHz, CDCl3) δ218.06,176.26,176.26,143.55,134.29,132.64,132.1 7,129.81(2C),128.95,123.50,122.22,120.49(2C),57.06,51.39,49.52,47 .77,47.24,47.05,43.86,37.08,35.43,34.49,32.62,29.44,28.75,27.36,26.86,26.70,25.94,25.66,24.20,21.18,21.08,20.18,19.74,17.54,15.98.

[0065] Electrospray ionization source-high resolution mass spectrometry (m / z): The calculated value of molecular protonated ion is 644.3624, and the measured value is 644.3602.

[0066] Example 3

[0067] The structural formula of a β-elemene acid derivative, specifically when the R1 group is 4-OCH3, is as follows:

[0068]

[0069] The preparation method of β-elemene acid derivative J3 includes the following steps:

[0070] S1. Add 1 mmol of β-elemeneic acid and 1.5 mmol of 4-(chloromethyl)-1-(4-methoxyphenyl)-1H-1,2,3-triazole to 5 mL of anhydrous acetonitrile, then add 2 mmol of anhydrous potassium carbonate (K2CO3), heat to 60 °C and stir for 12 h to obtain a mixed solution.

[0071] S2. Perform TCL detection on the mixed solution. After β-elemene acid has completely disappeared, pour the mixed solution into 10 mL of water, and then use 8 mL of dichloromethane (DCM) as the extractant to extract the mixed solution. Extract three times, and then wash with saturated sodium chloride and dry with anhydrous sodium sulfate to obtain the organic phase.

[0072] S3. The organic phase was purified by column chromatography by gradient elution with a mixture of petroleum ether and ethyl acetate in volume ratios of 20:1, 10:1 and 5:1 to obtain β-elemene acid derivative J3 in a yield of 49%.

[0073] The nuclear magnetic resonance (NMR) analysis of β-elemene acid derivative J3 yielded the following spectral data:

[0074] 1H NMR spectrum: 1 H NMR (300MHz, CDCl3) δ7.98 (s, 1H), 7.63 (t, J = 7.6Hz, 2H), 7.03 (d, J = 9.0Hz, 2H), 5.37 (d, J = 12.6Hz, 1H) ,5.26(d,J=12.7Hz,1H),5.04(t,J=7.1Hz,1H),3.88(s,3H),2.59–2.50(m,1H),2.47–2.45(m,1H),2.3 5(d,J=7.6Hz,1H),2.14–2.05(m,2H),1.97–1.81(m,7H),1.65(s,3H),1.77–1.58(m,6H),1.48(s,3H), 1.32–1.29(m,4H),1.22–1.20(m,1H),1.09(s,3H),1.05(s,3H),1.00(s,3H),0.88(s,3H),0.76(s,3H).

[0075] Carbon NMR spectrum: 13C NMR (75MHz, CDCl3) δ218.05,176.27,159.96,143.32,134.29,132.65,132.15, 130.32,123.51,122.36,122.13(2C),114.80(2C),57.13,55.66,51.40,49.52 ,47.77,47.25,47.04,43.86,37.09,35.47,34.50,32.62,29.70,29.44,28.74,27.37,26.86,26.68,25.93,25.66,24.21,21.09,20.17,19.75,17.55,15.98.

[0076] Electrospray ionization source-high resolution mass spectrometry (m / z): The calculated value of molecular protonated ion is 642.4265, and the measured value is 642.4271.

[0077] Example 4

[0078] The structural formula of a β-elemene acid derivative, specifically when the R1 group is 3,4,5-(OCH3)3, is as follows:

[0079]

[0080] The preparation method of β-elemene acid derivative J4 includes the following steps:

[0081] S1. Add 1 mmol of β-elemeneic acid and 1.5 mmol of 4-(chloromethyl)-1-(3,4,5-trimethoxyphenyl)-1H-1,2,3-triazole to 5 mL of anhydrous acetonitrile, then add 2 mmol of anhydrous potassium carbonate (K2CO3), heat to 60 °C and stir for 12 h to obtain a mixed solution.

[0082] S2. Perform TCL detection on the mixed solution. After β-elemene acid has completely disappeared, pour the mixed solution into 10 mL of water, and then use 8 mL of dichloromethane (DCM) as the extractant to extract the mixed solution. Extract three times, and then wash with saturated sodium chloride and dry with anhydrous sodium sulfate to obtain the organic phase.

[0083] S3. The organic phase was purified by column chromatography by gradient elution with a mixture of petroleum ether and ethyl acetate in volume ratios of 20:1, 10:1 and 5:1 to obtain β-elemene acid derivative J4 in a yield of 52%.

[0084] The nuclear magnetic resonance (NMR) analysis of β-elemene acid derivative J4 yielded the following spectral data:

[0085] 1H NMR spectrum: 1 H NMR (300MHz, CDCl3) δ8.04 (s, 1H), 6.94 (s, 2H), 5.31 (q, J = 12.7Hz, 2H), 5.04 (t, J = 6.8 Hz,1H),3.94(s,6H),3.90(s,3H),2.61–2.43(m,2H),2.39–2.30(m,1H),2.17–2.05(m, 2H),1.96-1.89(m,5H),1.85–1.77(m,2H),1.65(s,3H),1.62–1.50(m,6H),1.47(s,3H) ),1.34–1.25(m,5H),1.09(s,3H),1.05(s,3H),1.01(s,3H),0.88(s,3H),0.77(s,3H).

[0086] Carbon NMR spectrum: 13 C NMR(75MHz, CDCl3)δ217.89,176.40,153.98(2C),143.43,138.51,134.33,132.7 1,132.64,132.16,123.48,122.55,98.41(2C),61.08,57.10,56.46(2C),51.43,4 9.52, 47.73, 47.26, 47.04, 43.87, 37.10, 35.49, 34.49, 32.63, 29.43, 28.80, 27.38, 26.82, 26.64, 25.93, 25.66, 24.22, 21.20, 21.11, 20.16, 19.74, 17.54, 16.00.

[0087] Electrospray ionization source-high resolution mass spectrometry (m / z): The calculated value of molecular protonated ion is 702.4477, and the measured value is 702.4479.

[0088] Example 5

[0089] The structural formula of a β-elemene acid derivative, specifically when the R2 group is 4-H, is as follows:

[0090]

[0091] The preparation method of β-elemene acid derivative J5 includes the following steps:

[0092] S1. Add 1 mmol of β-elemeneic acid and 1.5 mmol of 4-(bromomethyl)-1-benzyl-1H-1,2,3-triazole to 5 mL of anhydrous acetonitrile, then add 2 mmol of anhydrous potassium carbonate (K2CO3), heat to 60 °C and stir for 12 h to obtain a mixed solution.

[0093] S2. Perform TCL detection on the mixed solution. After β-elemene acid has completely disappeared, pour the mixed solution into 10 mL of water, and then use 8 mL of dichloromethane (DCM) as the extractant to extract the mixed solution. Extract three times, and then wash with saturated sodium chloride and dry with anhydrous sodium sulfate to obtain the organic phase.

[0094] S3. The organic phase was purified by column chromatography by gradient elution with a mixture of petroleum ether and ethyl acetate in volume ratios of 20:1, 10:1 and 5:1 to obtain β-elemene acid derivative J5 in a yield of 45%.

[0095] The nuclear magnetic resonance (NMR) analysis of β-elemene acid derivative J5 yielded the following spectral data:

[0096] 1H NMR spectrum: 1 H NMR (300MHz, CDCl3) δ7.54(s,1H),7.37–7.32(m,3H),7.27–7.22(m,2H),5.53(s,J=4.1Hz,2H ),5.20(d,J=2.5Hz,2H),5.01(t,J=6.3Hz,1H),2.63–2.41(m,2H),2.29–2.27(m,1H),2.09–2 .06(m,2H),2.02–1.85(m,5H),1.76–1.72(m,2H),1.71(s,1H),1.66(s,3H),1.51(s,4H),1.4 7(s,3H),1.42-1.31(m,6H),1.10(s,3H),1.06(s,3H),1.03(s,3H),0.86(s,3H),0.75(s,3H).

[0097] Carbon NMR spectrum: 13C NMR (126MHz, CDCl3) δ218.03,176.21,143.26,134.41,134.29,132.71,132.07 ,129.15(2C),128.83,128.02(2C),123.85,123.53,57.20,54.19,51.46,49.53 ,47.66,47.27,46.98,43.86,37.12,35.48,34.53,32.57,29.43,28.71,27.38,26.81,26.67,25.85,25.67,24.20,21.16,21.12,20.18,19.77,17.54,15.96.

[0098] Electrospray ionization source-high resolution mass spectrometry (m / z): The calculated value of molecular protonated ion is 626.4316, and the measured value is 626.4318.

[0099] Example 6

[0100] The structural formula of a β-elemene acid derivative, specifically when the R2 group is 4-F, is as follows:

[0101]

[0102] The preparation method of β-elemene acid derivative J6 includes the following steps:

[0103] S1. Add 1 mmol of β-elemeneic acid and 1.5 mmol of 4-(bromomethyl)-1-(4-fluorobenzyl)-1H-1,2,3-triazole to 5 mL of anhydrous acetonitrile, then add 2 mmol of anhydrous potassium carbonate (K2CO3), heat to 60 °C and stir for 12 h to obtain a mixed solution.

[0104] S2. Perform TCL detection on the mixed solution. After β-elemene acid has completely disappeared, pour the mixed solution into 10 mL of water, and then use 8 mL of dichloromethane (DCM) as the extractant to extract the mixed solution. Extract three times, and then wash with saturated sodium chloride and dry with anhydrous sodium sulfate to obtain the organic phase.

[0105] S3. The organic phase was purified by column chromatography by gradient elution with a mixture of petroleum ether and ethyl acetate in volume ratios of 20:1, 10:1 and 5:1 to obtain β-elemene acid derivative J6 in a yield of 58%.

[0106] The nuclear magnetic resonance (NMR) analysis of β-elemene acid derivative J6 yielded the following spectral data:

[0107] 1H NMR spectrum:1 H NMR (300MHz, CDCl3) δ7.55 (s, 1H), 7.27–7.21 (t, 2H), 7.06 (t, J = 8.5Hz, 2H), 5.50 (s, 2 H),5.21(s,2H),5.01(t,J=6.6Hz,1H),2.63–2.45(m,2H),2.36–2.25(m,1H),2.17–2. 04(m,2H),2.03–1.88(m,5H),1.71(s,2H),1.66(s,3H),1.63–1.52(m,6H),1.47(s,3H ),1.41–1.22(m,5H),1.11(s,3H),1.07(s,3H),1.04(s,3H),0.87(s,3H),0.76(s,3H).

[0108] Carbon NMR spectrum: 13 C NMR(126MHz, CDCl3)δ217.90,176.21,162.88(d,J=248.4Hz),143.43,134.31,132.69,1 32.10,130.33(d,J=3.4Hz),129.92,129.85,123.78,123.49,116.25,116.08,57.17,53 .41,51.46,49.53,47.66,47.27,46.98,43.86,37.12,35.49,34.52,32.57,29.42,28.72,27.39,26.81,26.67,25.84,25.65,24.19,21.16,21.13,20.18,19.75,17.52,15.96.

[0109] Electrospray ionization source-high resolution mass spectrometry (m / z): The calculated value of molecular protonated ion is 644.4222, and the measured value is 644.4220.

[0110] Example 7

[0111] The structural formula of a β-elemene acid derivative, specifically when the R2 group is 4-Cl, is as follows:

[0112]

[0113] The preparation method of β-elemene acid derivative J7 includes the following steps:

[0114] S1. Add 1 mmol of β-elemeneic acid and 1.5 mmol of 4-(bromomethyl)-1-(4-chlorobenzyl)-1H-1,2,3-triazole to 5 mL of anhydrous acetonitrile, then add 2 mmol of anhydrous potassium carbonate (K2CO3), heat to 40 °C and stir for 24 h to obtain a mixed solution.

[0115] S2. Perform TCL detection on the mixed solution. After β-elemene acid has completely disappeared, pour the mixed solution into 10 mL of water, and then extract the mixed solution with 8 mL of dichloromethane (DCM) as the extractant. Then wash with saturated sodium chloride and dry with anhydrous sodium sulfate to obtain the organic phase.

[0116] S3. The organic phase was purified by column chromatography by gradient elution with a mixture of petroleum ether and ethyl acetate in volume ratios of 20:1, 10:1 and 5:1 to obtain β-elemene acid derivative J7 in a yield of 58%.

[0117] The nuclear magnetic resonance (NMR) analysis of β-elemene acid derivative J7 yielded the following spectral data:

[0118] 1H NMR spectrum: 1 H NMR (300MHz, CDCl3) δ7.56 (s, 1H), 7.35 (d, J = 8.4Hz, 2H), 7.20 (d, J = 8.4Hz, 2H), 5.50 (s, 2H) ,5.21(s,2H),5.01(t,J=7.1Hz,1H),2.64–2.45(m,2H),2.30–2.28(m,1H),2.17–2.05(m,2H ),2.03–1.88(m,5H),1.71(s,2H),1.67(s,3H),1.63–1.61(m,1H),1.58–1.49(m,5H),1.47( s,3H),1.34–1.26(m,5H),1.11(s,3H),1.07(s,3H),1.04(s,3H),0.87(s,3H),0.76(s,3H).

[0119] Carbon NMR spectrum: 13C NMR (75MHz, CDCl3) δ217.94,176.21,143.48,134.92,134.30,132.95,132.67, 132.12,129.37(2C),129.28(2C),123.87,123.47,57.15,53.40,51.44,49.52 ,47.65,47.26,46.98,43.85,37.11,35.48,34.53,32.57,29.42,28.72,27.38,26.80,26.66,25.84,25.67,24.19,21.17,21.13,20.17,19.75,17.52,15.96.

[0120] Electrospray ionization source-high resolution mass spectrometry (m / z): The calculated value of molecular protonated ion is 660.3927, and the measured value is 660.3932.

[0121] Example 8

[0122] The structural formula of a β-elemene acid derivative, specifically when the R2 group is 4-OCH3, is as follows:

[0123]

[0124] The preparation method of β-elemene acid derivative J8 includes the following steps:

[0125] S1. Add 1 mmol of β-elemeneic acid and 1.5 mmol of 4-(bromomethyl)-1-(4-methoxybenzyl)-1H-1,2,3-triazole to 5 mL of anhydrous acetonitrile, then add 2 mmol of anhydrous potassium carbonate (K2CO3), heat to 40 °C and stir for 24 h to obtain a mixed solution.

[0126] S2. Perform TCL detection on the mixed solution. After β-elemene acid has completely disappeared, pour the mixed solution into 10 mL of water, and then use 8 mL of dichloromethane (DCM) as the extractant to extract the mixed solution. Extract three times, and then wash with saturated sodium chloride and dry with anhydrous sodium sulfate to obtain the organic phase.

[0127] S3. The organic phase was purified by column chromatography by gradient elution with a mixture of petroleum ether and ethyl acetate in volume ratios of 20:1, 10:1 and 5:1 to obtain β-elemene acid derivative J8 in 55% yield.

[0128] The nuclear magnetic resonance (NMR) analysis of β-elemene acid derivative J8 yielded the following spectral data:

[0129] 1H NMR spectrum:1 H NMR (300MHz, CDCl3) δ7.51 (s, 1H), 7.23 (d, J = 8.4Hz, 2H), 6.90 (d, J = 8.4Hz, 2H), 5.46 (s, 2H), 5. 20(q,J=12.8Hz,2H),5.02(t,J=8.0Hz,1H),3.82(s,3H),2.62–2.45(m,2H),2.29(s,1H),2.06( d,J=8.1Hz,2H),1.99–1.75(m,5H),1.73(d,J=10.7Hz,3H),1.67(s,3H),1.62(s,2H),1.52(s,3 H),1.48(s,3H),1.29(s,5H),1.11(s,3H),1.07(s,3H),1.04(s,3H),0.87(s,3H),0.76(s,3H).

[0130] Carbon NMR spectrum: 13 C NMR (75MHz, CDCl3) δ217.98,176.20,172.83,159.97,143.14,134.27,132.70,13 2.07,129.60(2C),126.35,123.63,123.53,114.49(2C),57.22,55.31,53.73,51 .44,49.52,47.66,47.26,46.97,43.84,37.11,35.48,34.53,32.56,29.43,28.68,27.37,26.81,26.66,25.85,25.67,24.19,21.12,20.18,19.75,17.53,15.95.

[0131] Electrospray ionization source-high resolution mass spectrometry (m / z): The calculated value of molecular protonated ion is 656.4422, and the measured value is 656.4414.

[0132] Test 1

[0133] The inhibitory rates of β-elemene derivatives (β-elemene derivative J1, β-elemene derivative J2, β-elemene derivative J3, β-elemene derivative J4, β-elemene derivative J5, β-elemene derivative J6, β-elemene derivative J7, and β-elemene derivative J8) and β-elemene acid on human cervical cancer cells SiHa, human liver cancer cells HepG2, human colon cancer cells HCT-116, and human lung cancer cells A549 were determined by the MTT assay. The results are shown in Table 1. Cells were then divided into groups of approximately 4 × 10⁻⁶ cells. 3Cells were evenly seeded in 96-well plates and incubated at 37°C with 5% CO2 for 12 hours. After cell attachment, the cells were treated with the drug and cultured for another 72 hours. The supernatant was discarded, and 150 μL of serum-free medium-concentrated MTT solution (0.5 mg / mL, freshly prepared) was added to each well, and the cells were incubated for another 4 hours. The supernatant was then discarded, and 150 μL of DMSO was added to fully dissolve formazan (protected from light, shaken for 10 min), and the OD value at 490 nm was measured. Using the DMSO group as a blank control, the inhibition rate of β-elemene derivatives on different cell types was calculated using the following formula:

[0134] Inhibition rate (%) = (1 - OD value of dosing well / OD value of control well) × 100, where half-maximal inhibitory concentration (IC50) is the concentration at half maximum. 50 The result was obtained by plotting the drug concentration and inhibition rate curves.

[0135] Table 1. Effects of β-elemene and its derivatives on IC50 in different cancer cells. 50

[0136]

[0137]

[0138] Table 1 shows that for SiHa cells: the IC50 values ​​for β-elemene derivative J1 and β-elemene derivative J3 are... 50 The concentrations of β-elemene derivative J1 and β-elemene acid were 0.96 μM and 9.64 μM, respectively, showing strong anti-proliferative activity. The activity of β-elemene acid derivative J1 was 56.7 times stronger than that of the lead compound (β-elemene acid). For HepG2 cells: β-elemene acid derivative J1 exhibited the strongest anti-HepG2 cell proliferation activity, with an IC50 concentration of 0.96 μM and 9.64 μM, respectively. 50 The concentration was 2.51 μM, 24.5 times that of the lead compound. For HCT-116 cells: the β-elemene derivative J1 exhibited the strongest activity, with an IC50 concentration of 2.51 μM. 50 The concentration was 1.10 μM, which was 42.6 times that of the lead compound. In A549 cells: the β-elemene acid derivative J1 exhibited the strongest activity, with an IC50 concentration of 1.10 μM. 50 The concentration was 3.18 μM, which is 15.7 times that of the lead compound. Therefore, the β-elemene derivative J1 showed significantly stronger inhibitory effects on SiHa, HepG2, HCT-116, and A549 cells than β-elemene, indicating its potential as an antitumor drug. Furthermore, it can be combined with pharmaceutically acceptable carriers or excipients to prepare β-elemene derivative compositions.

[0139] Test 2

[0140] Toxicity to normal cells is one of the major obstacles to the drug development of antitumor compounds, and most anticancer drugs have a narrow therapeutic index. Therefore, evaluating the selectivity index (SI) of compounds against normal and tumor cells is essential in anticancer drug research. The selectivity index SI = IC 50 (Normal cells) / IC 50 (Tumor cells). To investigate whether the enhanced antiproliferative effect of β-elemene derivative J1 on tumor cells is due to cytotoxicity, a cytotoxicity assay was performed on normal human hepatocytes L02. As shown in Table 2, β-elemene derivative J1 showed low toxicity to normal human hepatocytes, with an IC50 value of [missing value]. 50 The concentration was 49.87 μM, significantly lower than that of β-elemene acid. β-elemene acid derivative J1 exhibited high selectivity for all tested tumor cells (SiHa cells, HepG2 cells, HCT-116 cells, and A549 cells), with an SI of 51.95 for SiHa cells and 45.34 for HCT-116 cells, demonstrating a significantly stronger selectivity coefficient than β-elemene acid.

[0141] Table 2 Selectivity index of β-elemene derivative J1 and β-elemene acid

[0142] Selectivity index SI = IC 50 (Normal cells) / IC 50 (Tumor cells)

[0143] Therefore, the present invention adopts the above-mentioned β-elemene acid derivative and its preparation method and application, using tetracyclic triterpenoid β-elemene acid as a lead compound, and introducing a 1,2,3-triazole structure with anticancer activity. The generated β-elemene acid derivative has low toxicity to normal human liver cells, high selectivity for tumor cells, and significant anti-proliferation effect.

[0144] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A β-elemene acid derivative, characterized in that, The structural formula of the β-elemene acid derivative is: The structural formula of the R-based base is: One of the following: R1 group is one of 4-H, 4-Cl, 4-OCH3 and 3,4,5-(OCH3)3; R2 group is one of 4-H, 4-F, 4-Cl and 4-OCH3.

2. The β-elemene acid derivative according to claim 1, characterized in that, When the R1 group is 4-H, the structural formula of the β-elemene acid derivative is: When the R1 group is 4-Cl, the structural formula of the β-elemene acid derivative is: When the R1 group is 4-OCH3, the structural formula of the β-elemene acid derivative is: When the R1 group is 3,4,5-(OCH3)3, the structural formula of the β-elemene acid derivative is:

3. The β-elemene acid derivative according to claim 1, characterized in that, When the R2 group is 4-H, the structural formula of the β-elemene acid derivative is: When the R2 group is 4-F, the structural formula of the β-elemene acid derivative is: When the R2 group is 4-Cl, the structural formula of the β-elemene acid derivative is: When the R2 group is 4-OCH3, the structural formula of the β-elemene acid derivative is:

4. A method for preparing a β-elemene acid derivative as described in any one of claims 1-3, characterized in that, Includes the following steps: S1. Add β-elemene acid and alkyl bromide / chlorine derivative to anhydrous acetonitrile, then add anhydrous potassium carbonate, heat and stir to obtain a mixed solution; S2. The mixed solution was extracted using dichloromethane as the extractant, and the organic phase was obtained after washing and drying. S3. The organic phase was purified by column chromatography to obtain β-elemene acid derivative.

5. The method for preparing a β-elemene acid derivative according to claim 4, characterized in that, The alkyl bromide / chlorine derivative is one of 4-(chloromethyl)-1-phenyl-1H-1,2,3-triazole, 4-(chloromethyl)-1-(4-chlorophenyl)-1H-1,2,3-triazole, 4-(chloromethyl)-1-(4-methoxyphenyl)-1H-1,2,3-triazole, 4-(chloromethyl)-1-(3,4,5-trimethoxyphenyl)-1H-1,2,3-triazole, 4-(bromomethyl)-1-benzyl-1H-1,2,3-triazole, 4-(bromomethyl)-1-(4-fluorobenzyl)-1H-1,2,3-triazole, 4-(bromomethyl)-1-(4-chlorobenzyl)-1H-1,2,3-triazole, and 4-(bromomethyl)-1-(4-methoxybenzyl)-1H-1,2,3-triazole.

6. The method for preparing a β-elemene acid derivative according to claim 4, characterized in that, The molar ratio of β-elemene, alkyl bromide / chloride derivatives and anhydrous potassium carbonate in S1 is 1:(1.5-2):(2-3).

7. The method for preparing a β-elemene acid derivative according to claim 4, characterized in that, In S1, the heating temperature is 40℃-60℃, the heating time is 12h-24h, and the stirring time is 12h-24h.

8. The method for preparing a β-elemene acid derivative according to claim 4, characterized in that, In S3, the eluent used for column chromatography purification is a mixed solution of petroleum ether and ethyl acetate; the volume ratio of the petroleum ether to the ethyl acetate is (5-20):

1.

9. The application of a β-elemene acid derivative, characterized in that, The β-elemene acid derivative according to any one of claims 1-3 is used in the preparation of antitumor drugs.

10. A β-elemene acid derivative composition, characterized in that, It includes a β-elemene acid derivative as described in any one of claims 1-3, and a pharmaceutically acceptable pharmaceutical carrier or excipient.

Images