Antitumor covalently modified curcumol derivatives and their preparation methods and applications

Abstract

The present invention belongs to the field of pharmaceutical technology, and particularly relates to curcumol derivatives containing Michael receptors, their synthesis methods and their applications in the treatment of malignant tumors; with covalent modification as the core strategy, we synthesized two types of curcumol derivatives with reactive Michael receptors. In the structures of these derivatives, there are reactive Michael receptors such as α,β-unsaturated aldehydes, α,β-unsaturated acids, α,β-unsaturated amides, α,β-unsaturated ketones, and α,β-unsaturated nitriles, which exhibit better inhibitory activities against various tumor cells than curcumol, and are expected to be developed into highly effective and low-toxic anti-tumor drugs.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and specifically relates to curcumol derivatives containing Michael receptors, their synthesis methods and their applications in the treatment of malignant tumors. Background Art

[0002] Due to factors such as environmental pollution and changes in human lifestyle, the incidence of malignant tumors has been increasing annually, seriously threatening human health. The latest statistical data released by the World Health Organization shows that in 2020, the number of newly diagnosed cases of malignant tumors globally was 19.3 million, and the number of newly deceased cases was 10 million; among which, the number of newly diagnosed cases in China was approximately 4.57 million, and the number of deceased cases was approximately 3 million, both ranking first in the world.

[0003] Small molecule chemical drugs are the main means to combat malignant tumors. The commonly used anti-tumor drugs clinically mainly include cytotoxic drugs and targeted drugs. However, cytotoxic drugs usually show strong toxic and side effects, seriously affecting the quality of life of malignant tumor patients. Targeted drugs show increasing drug resistance, becoming a bottleneck in their clinical application. Therefore, there is an urgent need to develop anti-tumor drugs with high efficiency, low toxicity and high therapeutic index to address this challenge.

[0004] Curcuma zedoaria is the dried rhizome of Curcuma phaeocaulis Val., Curcuma kwangsiensis S.G. Lee et C.F. Liang, Curcuma wenyujin Y.H. Chen et C.Ling of the Zingiberaceae family. It is a traditional genuine medicinal material with a long history of use in China. Curcuma zedoaria was first recorded in "Yaoxinglun" by Zhen Quan in the Tang Dynasty. "Compendium of Materia Medica" records that "Curcuma zedoaria is pungent, warm and non-toxic, its color is black, and it can break the blood in qi." "Explanation of the Compendium of Materia Medica" once recorded that "Curcuma phaeocaulis tastes pungent and has a strong nature, specializing in breaking the blood in qi, mainly for dissipating stasis and resolving masses, removing accumulations and lumps, amenorrhea and blood stasis, and traumatic pain." "Cui Jinqiu Bencaoshu Lv" also pointed out that "Guangmao is Curcuma zedoaria, and it is used for promoting qi and breaking blood, dissipating stasis and resolving masses." Correspondingly, modern medicine has proved that the volatile oil of Curcuma zedoaria has various therapeutic functions such as anti-tumor, antibacterial, anti-early pregnancy, anti-inflammatory, and liver protection. Especially the anti-tumor effect is definite and obvious, which has attracted wide attention. Curcumol is one of the material bases for the anti-tumor effect of the volatile oil of Curcuma zedoaria. It can play an anti-tumor role through various mechanisms such as blocking the cell cycle and inducing apoptosis, and has inhibitory activity against various types of malignant tumor cells such as colon cancer, bladder cancer, melanoma, liver cancer, osteosarcoma, nasopharyngeal carcinoma, breast cancer, ovarian cancer, cervical cancer, lung cancer, gastric cancer, and cholangiocarcinoma. Moreover, as an external medicine, it was once briefly applied to the clinical treatment of cervical cancer in China. However, problems such as poor anti-tumor activity, poor water solubility, and unclear mechanism of action have severely restricted the further development of new drugs. Summary of the Invention

[0005] To solve the deficiencies of the prior art, the purpose of the present invention is to prepare curcumol derivatives containing Michael receptors according to the covalent modification strategy and study their anti-tumor activities.

[0006] Covalent modification refers to introducing highly reactive reactive groups into bioactive molecules, making them react with endogenous target proteins to form covalent bonds, irreversibly and continuously changing the functions of target proteins, and then exerting therapeutic effects. Since it can make the binding of bioactive molecules to target proteins stronger and the action time longer, this strategy has become an effective way to enhance drug efficacy, overcome acquired drug resistance, and develop new anti-tumor drugs.

[0007] In view of this, the first aspect of the present invention discloses a curcumol derivative containing a reactive Michael receptor.

[0008] Preferably, the general structural formula of the above curcumol derivative is as follows:

[0009]

[0010] R in the general structural formula I 1The group represents a hydrogen atom, an alkyl group, an aryl group, a heteroaryl group, a heteroarylmethyl group, etc.; R 2 The group represents a hydrogen atom, an alkyl group, etc. Among them:

[0011] The alkyl group is a C1-C6 alkyl group or a deuterated alkyl group;

[0012] The aryl group is a phenyl group, a substituted phenyl group;

[0013] For the substituted phenyl group, the substituent is a halogen;

[0014] The heteroaryl group is a pyridyl group or a substituted pyridyl group;

[0015] The heteroarylmethyl group is a pyridylmethyl group, a substituted pyridylmethyl group.

[0016]

[0017] R in the general structural formula II 3 The group represents a hydrogen atom, a hydroxyl group, a cyano group, a nitrogen-containing heterocycle. Among them: the nitrogen-containing heterocycle is a 4- to 7-membered nitrogen-containing heterocycle.

[0018]

[0019] R in the general structural formula III 4 and R 5 The group represents an alkyl group. Among them:

[0020] The alkyl group is a C1-C6 alkyl group or a deuterated alkyl group.

[0021] Further preferably, the derivative is the following compound:

[0022]

[0023]

[0024]

[0025]

[0026] The two types of curcumol derivatives described in the present invention are obtained through the following preparation method:

[0027]

[0028] The second aspect of the present invention also provides a preparation method for the above-mentioned derivative, and this method includes the following steps:

[0029] Curcumol first reacts with mCPBA to generate an epoxy intermediate, and then under heating and alkaline conditions, after the epoxide ring opens, elimination occurs to obtain the hydroxy compound II, with a yield of 56%.

[0030] Compound II was oxidized after reacting with Dess-Martin oxidant to obtain Compound S1 in a yield of 78%.

[0031] Compound S1 was oxidized under the action of sodium chlorite and hydrogen peroxide to obtain Compound S2 in a yield of 93%.

[0032] Compound S2 underwent condensation reactions with various amines to obtain Compounds S3 - S17 in yields of 86 - 97%.

[0033]

[0034] Under room temperature conditions, with ruthenium trichloride hydrate as a catalyst, curcumol was oxidized by sodium periodate to obtain Compound III in a yield of 85%.

[0035] Compound III underwent aldol condensation reaction with paraformaldehyde to obtain Compound S18 in a yield of 37%.

[0036] Compound S18 underwent addition reaction with dimethylamine to obtain Compound S19 in a yield of 98%.

[0037] The third aspect of the present invention discloses the application of the above-mentioned curcumol derivatives in the preparation of anti-cancer drugs. The active ingredient in the drug is mainly the above-mentioned curcumol derivatives. The cancers include but are not limited to various types of malignant tumor cancers such as colon cancer, bladder cancer, melanoma, liver cancer, osteosarcoma, nasopharyngeal cancer, breast cancer, ovarian cancer, cervical cancer, lung cancer, gastric cancer, cholangiocarcinoma, etc.

[0038] Compared with the prior art, the present invention has the following remarkable effects:

[0039] With covalent modification as the core strategy, the present invention synthesized two types of curcumol derivatives with reactive Michael acceptors. In the structures of these derivatives, there are reactive Michael acceptors such as α,β-unsaturated aldehyde, α,β-unsaturated acid, α,β-unsaturated amide, α,β-unsaturated ketone, α,β-unsaturated nitrile, etc., which show better inhibitory activities against various tumor cells than curcumol, and are expected to be developed into anti-tumor drugs with high efficiency and low toxicity. Specific Embodiments

[0040] The present invention will be further illustrated by the following examples, but the present invention is not limited to the scope of the described examples. For the experimental methods without specific conditions in the following examples, they are carried out according to conventional methods and conditions, or selected according to the product instructions.

[0041] Example 1: Synthesis of Compound S1:

[0042] At 0 °C, NaHCO3 (133 mg, 1.58 mmol, 2 equiv.) and Dess-Martin oxidant (503 mg, 1.19 mmol, 1.5 equiv.) were added to a solution of compound II (200 mg, 0.79 mmol, 1 equiv.) in dichloromethane (15 mL). The mixture was stirred at room temperature for 6 h. After completion of the reaction, it was quenched with saturated NH4Cl solution. The mixture was extracted with ethyl acetate, the organic layer was separated, dried over anhydrous sodium sulfate, concentrated, rotary evaporated and purified by column chromatography (PE:EA = 5:1) to obtain compound S1 as a white solid (20 mg, yield 78%). 1 1H NMR (400 MHz, CDCl3) δ 9.54 (s, 1H), 6.81 (s, 1H), 3.02 (s, 1H), 2.39–2.23 (m, 2H), 2.20–2.08 (m, 1H), 2.00–1.85 (m, 2H), 1.75 (m, 1H), 1.48–1.36 (m, 2H), 1.26 (m, 2H), 1.06 (d, J = 6.5 Hz, 3H), 1.03 (d, J = 6.4 Hz, 3H), 0.90 (d, J = 6.5 Hz, 3H). HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 15 H 23 O3 251.1647; Found 251.1645.

[0043] Example 2: Synthesis of compound S2

[0044] At room temperature, NaClO2 (100 mg, 1.11 mmol, 1.4 equiv.) was added to a mixed solution of compound S1 (200 mg, 0.79 mmol, 1 equiv.), NaH2PO4 (28 mg, 0.24 mmol, 0.3 equiv.) and 30% aqueous H2O2 (0.08 mL) in CH3CN (0.6 mL) and water (0.3 mL). The reaction mixture was stirred at room temperature overnight. After completion of the reaction, it was acidified with 2N aqueous HCl and then filtered to obtain the desired compound S2 as a white solid (25 mg, yield 93%). 11H NMR (400 MHz, DMSO-d6) δ 6.58 (s, 1H), 5.82 (s, 1H), 2.30 (t, J = 8.2 Hz, 1H), 2.12 (t, J = 11.9 Hz, 1H), 1.85 (m, 2H), 1.65 (m, 2H), 1.41 (m, 2H), 1.33 (m, 1H), 1.28–1.20 (m, 1H), 0.94 (d, J = 6.4 Hz, 3H), 0.90 (d, J = 6.4 Hz, 3H), 0.85 (d, J = 6.5 Hz, 3H). HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 15 H 23 O4 267.1596; Found 267.1595.

[0045] Example 3: Synthesis of Compound S3

[0046] At room temperature, piperazine (7.6 mg, 0.088 mmol, 1.1 equiv.) was added to a solution of compound S2 (20 mg, 0.08 mmol, 1 equiv.), EDCI (18 mg, 0.096 mmol, 1.2 equiv.), HOBt (13 mg, 0.096 mmol, 1.2 equiv.), and DIPEA (42 μL, 0.24 mmol, 3 equiv.) in anhydrous DCM. The mixture was stirred overnight at 25 °C. After completion of the reaction, the mixture was extracted three times with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, concentrated, rotary evaporated, and purified by column chromatography to obtain compound S3 as a white solid (29 mg, 86% yield). 1 1H NMR (400 MHz, CDCl3) δ 5.91 (s, 1H), 3.60 (m, 4H), 2.48 (t, J = 9.2 Hz, 1H), 2.24 (m, 1H), 1.95–1.77 (m, 6H), 1.70 (m, 2H), 1.61–1.54 (m, 1H), 1.51–1.36 (m, 4H), 1.02 (d, J = 6.1 Hz, 3H), 0.98 (d, J = 6.5 Hz, 3H), 0.88 (d, J = 6.5 Hz, 3H). HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 19 H 31 N2O3 333.2335; Found 333.2337.

[0047] Example 4: Synthesis of Compound S4

[0048] Piperazine was replaced with N-methylpiperazine at the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, obtaining 29 mg of white solid S4 with a yield of 89%. 1 HNMR(400MHz,CDCl3)δ5.87(d,J=1.5Hz,1H),3.80(m,4H),2.80(s,3H),2.48(t,J=9.2Hz,1H),2.25(m,2H),1.99–1.79(m,3H),1.78–1.59(m,5H),1.55–1.48(m,2H),1.42(m,1H),1.02(d,J=6.3Hz,3H),0.98(d,J=6.4Hz,3H),0.88(d,J=6.5Hz,3H).HRMS(ESI–TOF)m / z:[M+H] + Calcd for C 20 H 33 N2O3349.2491;Found 349.2489.

[0049] Example 5: Synthesis of Compound S5

[0050] Piperazine was replaced with morpholine at the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, obtaining 31 mg of white solid S5 with a yield of 93%. 1 H NMR(400MHz,CDCl3)δ5.87(d,J=1.6Hz,1H),3.90–3.46(m,8H),2.93(s,1H),2.50(m,1H),2.25(m,1H)1.99–1.81(m,3H),1.64–1.57(m,2H),1.50(m,2H),1.42(m,1H),1.02(d,J=6.3Hz,3H),0.98(d,J=6.4Hz,3H),0.88(d,J=6.4Hz,3H).HRMS(ESI–TOF)m / z:[M+H] + Calcd for C 19 H 30 NO4336.2175;Found 336.2173.

[0051] Example 6: Synthesis of Compound S6

[0052] Piperazine was replaced with piperidine at the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, obtaining 32 mg of white solid S4 with a yield of 97%. 11H NMR (400 MHz, CDCl3) δ 5.84 (d, J = 1.6 Hz, 1H), 3.86–3.38 (m, 4H), 3.00 (s, 1H), 2.51–2.42 (m, 1H), 2.24 (m, 1H), 1.97–1.80 (m, 3H), 1.71–1.48 (m, 10H), 1.42 (m, 1H), 1.02 (d, J = 6.2 Hz, 3H), 0.98 (d, J = 6.4 Hz, 3H), 0.88 (d, J = 6.5 Hz, 3H). HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 20 H 32 NO3 334.4800; Found 334.4801.

[0053] Example 7: Synthesis of Compound S7

[0054] Piperazine was replaced with pyrrolidine under the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, obtaining 30 mg of white solid S4 with a yield of 93%. 1 1H NMR (400 MHz, CDCl3) δ 6.04–6.00 (s, 1H), 3.73–3.39 (m, 4H), 3.07 (s, 1H), 2.53 (t, J = 8.7 Hz, 1H), 2.24 (m, 1H), 1.99–1.80 (m, 7H), 1.68–1.46 (m, 4H), 1.40 (m, 1H), 1.02 (d, J = 6.0 Hz, 3H), 0.98 (d, J = 6.4 Hz, 3H), 0.88 (d, J = 6.5 Hz, 3H). HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 19 H 30 NO3 320.2226; Found 320.2228.

[0055] Example 8: Synthesis of Compound S8

[0056] Piperazine was replaced with azetidine under the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, obtaining 29 mg of white solid S4 with a yield of 96%. 1HNMR (400MHz, CDCl3) δ6.10 (d, J = 1.4Hz, 1H), 4.38–3.98 (m, 4H), 2.85 (s, 1H), 2.47 (m, 1H), 2.34–2.18 (m, 3H), 2.06–1.81 (m, 3H), 1.69–1. 55(m,2H),1.52–1.41(m,2H),1.34(m,1H),1.01(d,J=6.7Hz,3H),0.99(d,J=6.5Hz,3H),0.88(d,J=6.5Hz,3H).HRMS(ESI–TOF)m / z:[M+H] + Calcdfor C 18 H 28 NO3306.4260;Found 306.4261.

[0057] Example 9: Synthesis of Compound S9

[0058] Piperazine was replaced by diethylamine at the same equivalent ratio. The remaining reagents and operations were the same as those for the synthesis of S3 to obtain 31 mg of white solid S4 with a yield of 97%. 1 HNMR(400MHz, CDCl3)δ5.86(s,1H),3.40(m,4H),2.88(s,1H),2.44(t,J=8.6Hz,1H),2.25(m,1H),1.93–1.80(m,2H),1.69–1.51(m,4 H),1.41(m,1H),1.15(t,J=7.1Hz,6H),1.02(d,J=6.0Hz,3H),0.99(d,J=6.4Hz,3H),0.89(d,J=6.4Hz,3H).HRMS(ESI–TOF)m / z:[M+H] + C 19 H 32 NO3322.2382;Found 322.2382.

[0059] Example 10: Synthesis of Compound S10

[0060] Piperazine was replaced by n-propylamine at the same equivalent ratio. The remaining reagents and operations were the same as those for the synthesis of S3 to obtain 30 mg of white solid S4 with a yield of 97%. 11H NMR (400 MHz, CDCl3) δ 6.47 (d, J = 1.5 Hz, 1H), 5.79 (s, 1H), 3.29 (m, 2H), 2.93 (s, 1H), 2.46–2.37 (m, 1H), 2.23 (m, 1H), 2.08–1.82 (m, 3H), 1.72–1.46 (m, 7H), 1.31 (m, 1H), 1.02 (m, 5H), 0.94 (t, J = 7.4 Hz, 3H), 0.87 (d, J = 6.5 Hz, 3H). 13 13C NMR (150 MHz, CDCl3) δ 166.9, 140.0, 132.2, 103.2, 87.4, 60.2, 48.0, 41.3, 40.4, 36.0, 31.2, 30.5, 28.1, 22.8, 22.6, 21.5, 11.5, 11.4. HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 18 H 30 NO3 308.2226; Found 308.2225.

[0061] Example 11: Synthesis of Compound S11

[0062] Piperazine was replaced with n-butylamine at the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, to obtain 30 mg of white solid S4 with a yield of 92%. 1 1H NMR (400 MHz, CDCl3) δ 6.46 (d, J = 1.4 Hz, 1H), 5.74 (s, 1H), 3.32 (m, 2H), 2.81 (s, 1H), 2.42 (m, 1H), 2.23 (m, 1H), 2.03–1.84 (m, 3H), 1.70–1.61 (m, 2H), 1.56–1.46 (m, 4H), 1.41–1.26 (m, 4H), 1.02 (m, 5H), 0.94 (t, J = 7.3 Hz, 3H), 0.88 (d, J = 6.5 Hz, 3H). HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 19 H 32 NO3 322.4690; Found 322.4693.

[0063] Example 12: Synthesis of Compound S12

[0064] Piperazine was replaced with aniline at the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, to obtain 32 mg of white solid S4 with a yield of 95%. 11H NMR (400 MHz, CDCl3) δ 7.56 (d, J = 8.1 Hz, 2H), 7.34 (t, J = 7.8 Hz, 2H), 7.12 (t, J = 7.4 Hz, 1H), 6.62 (d, J = 1.5 Hz, 1H), 3.05 (s, 1H), 2.56 (t, J = 9.0 Hz, 1H), 2.27 (m, 1H), 2.12–2.01 (m, 1H), 2.01–1.85 (m, 2H), 1.70 (m, 2H), 1.57–1.46 (m, 2H), 1.40–1.31 (m, 1H), 1.04 (t, J = 5.8 Hz, 6H), 0.89 (d, J = 6.5 Hz, 3H). HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 18 H 30 NO3 342.2069; Found 342.2070.

[0065] Example 13: Synthesis of Compound S13

[0066] Piperazine was replaced with N-methylaniline in the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, obtaining 32 mg of white solid S4 with a yield of 90%. 1 1H NMR (400 MHz, CDCl3) δ 7.37 (m, 2H), 7.31–7.28 (m, 1H), 7.19–7.10 (m, 2H), 5.76 (d, J = 1.5 Hz, 1H), 3.33 (s, 3H). 2.05 (s, 1H), 1.95–1.83 (m, 2H), 1.80–1.70 (m, 2H), 1.53–1.47 (m, 3H), 1.28–1.18 (m, 3H), 0.94 (d, J = 6.6 Hz, 3H), 0.79 (d, J = 4.2 Hz, 3H), 0.70 (d, J = 2.4 Hz, 3H). HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 22 H 30 NO3 356.2226; Found 356.2225.

[0067] Example 14: Synthesis of Compound S14

[0068] Piperazine was replaced with 4-fluoroaniline in the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, obtaining 33 mg of white solid S4 with a yield of 91%. 1HNMR(400MHz, CDCl3) δ 7.65(s, 1H), 7.52(m, 2H), 7.02(t, J = 8.5Hz, 2H), 6.62(s, 1H), 2.54(t, J = 9.0Hz, 1H), 2.27(t, J = 11.9Hz, 1H), 2.07(t, J = 9.4Hz, 1H), 1.95(m, 2H), 1.76–1.46(m, 4H), 1.35(m, 1H), 1.04(t, J = 5.9Hz, 6H), 0.89(d, J = 6.6Hz, 3H). HRMS(ESI–TOF) m / z: [M + H] + C 21 H 27 FNO3 360.1975; Found 360.1974.

[0069] Example 15: Synthesis of Compound S15

[0070] Piperazine was replaced with 2-aminopyridine at the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, obtaining 33 mg of white solid S4 with a yield of 95%. 1 HNMR(400MHz, CDCl3) δ 8.74(dd, J = 4.5, 1.4Hz, 1H), 8.44(dd, J = 8.4, 1.4Hz, 1H), 7.50(d, J = 1.5Hz, 1H), 7.46(dd, J = 8.4, 4.5Hz, 1H), 2.53(m, 1H), 2.34(m, 1H), 2.23(m, 1H), 2.01–1.91(m, 2H), 1.87–1.63(m, 5H), 1.42(m, 1H), 1.09(d, J = 6.5Hz, 3H), 1.06(d, J = 6.2Hz, 3H), 0.95(d, J = 6.5Hz, 3H). HRMS(ESI–TOF) m / z: [M + H] + Calcd for C 20 H 27 N2O3 343.2022; Found 343.2023.

[0071] Example 16: Synthesis of Compound S16

[0072] Piperazine was replaced with 4-aminopyridine at the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, obtaining 31 mg of white solid S4 with a yield of 92%. 11H NMR (400 MHz, CDCl3) δ 8.54–8.44 (m, 2H), 8.01 (s, 1H), 7.58–7.52 (m, 2H), 6.69 (d, J = 1.4 Hz, 1H), 2.59–2.53 (m, 1H), 2.28 (m, 1H), 2.11–1.88 (m, 3H), 1.74 (m, 1H), 1.67–1.59 (m, 1H), 1.56–1.45 (m, 2H), 1.35 (m, 1H), 1.07–1.00 (m, 6H), 0.89 (d, J = 6.5 Hz, 3H). HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 20 H 27 N2O3 343.2022; Found 343.2024.

[0073] Example 17: Synthesis of Compound S17

[0074] Piperazine was replaced with 4-methylaminopyridine in the same equivalent ratio, and the remaining reagents and operations were the same as those in the synthesis of S3, to obtain 34 mg of white solid S4 with a yield of 97%. 1 1H NMR (400 MHz, CDCl3) δ 8.56 (d, J = 5.1 Hz, 1H), 7.70 (m, 1H), 7.30 (d, J = 9.7 Hz, 2H), 6.69 (d, J = 1.6 Hz, 1H), 4.64 (t, J = 4.1 Hz, 2H), 2.47 (t, J = 9.1 Hz, 1H), 2.25 (m, 1H), 2.05 (s, 1H), 1.91 (m, 2H), 1.76–1.67 (m, 1H), 1.54–1.47 (m, 2H), 1.33 (m, 2H), 1.03 (m, 6H), 0.87 (d, J = 6.8 Hz, 3H). HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 21 H 29 N2O3 357.2178; Found 357.2175.

[0075] Example 18: Synthesis of Compound S18

[0076] At room temperature, paraformaldehyde (63 mg, 2.1 mmol, 5 equiv.), piperidine (6 μL, 0.055 mmol, 0.13 equiv.) and AcOH (5 μL, 0.092 mmol, 0.22 equiv.) were successively added to a solution of compound III (100 mg, 0.42 mmol, 1 equiv.) in DMF (0.5 mL). The resulting mixture was heated at 90 °C for 1 hour. After completion of the reaction, water was added to the reaction solution, and the mixture was extracted three times with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, concentrated, rotary evaporated and purified by column chromatography (PE:EA = 5:1) to obtain 10 mg of white solid S18 with a yield of 37%. 1 HNMR (400 MHz, CDCl3) δ 6.28 (d, J = 1.6 Hz, 1H), 5.77 (d, J = 1.6 Hz, 1H), 2.46 (m, 1H), 2.30 (m, 1H), 2.07–1.96 (m, 2H), 1.88–1.77 (m, 2H), 1.76–1.67 (m, 1H), 1.55–1.45 (m, 1H), 1.37 (m, 1H), 1.24 (m, 1H), 1.01 (d, J = 6.5 Hz, 3H), 0.89 (d, J = 6.4 Hz, 3H), 0.85 (d, J = 6.6 Hz, 3H). 13 C NMR (150 MHz, CDCl3) δ 200.7, 144.7, 121.9, 105.5, 86.2, 60.0, 56.5, 40.9, 35.4, 31.0, 29.8, 27.2, 22.4, 20.6, 11.6. HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 15 H 23 O3 251.1647; Found 251.1645.

[0077] Example 19: Synthesis of compound S19

[0078] Dimethylamine (2 M in THF, 45 μL, 0.09 mmol, 1.1 equiv.) was slowly added to a solution of compound S18 (20 mg, 0.08 mmol, 1 equiv.) in ethanol (0.5 mL). The reaction solution was stirred at room temperature for 24 hours. After completion of the reaction, the solvent was removed under reduced pressure and purified by column chromatography (PE:EA = 2:1) to obtain 29 mg of yellow oily compound S19 with a yield of 98%. 11H NMR (400 MHz, CDCl3) δ 2.72 (m, 2H), 2.62–2.52 (m, 4H), 2.48–2.38 (m, 2H), 2.16 (s, 6H), 2.09 (m, 2H), 2.04–1.93 (m, 3H), 1.89–1.78 (m, 2H), 0.96 (d, J = 6.8 Hz, 3H), 0.92 (d, J = 6.8 Hz, 3H), 0.84 (d, J = 6.8 Hz, 3H). HRMS (ESI–TOF) m / z: [M+H] + Calcd for C 18 H 30 NO3 296.2226; Found 296.2228.

[0079] The covalently modified curcumol derivatives synthesized in this invention have antitumor activity, and the results of the cell experiments are as follows:

[0080] The corresponding cell proliferation inhibition activity analysis was carried out using the CCK-8 method, and the specific operations are as follows:

[0081] 1. Sample preparation: After dissolving with DMSO (Merck), add PBS (-) to make a 0.6 mM solution, and then dilute it to 30 μM or 10 μM solution with PBS (-) containing DMSO.

[0082] 2. Tumor cell lines: U87, A172.

[0083] 3. Culture medium: RPMI 1640 + 10% FBS + double antibodies

[0084] 4. Other materials: Full wavelength multifunctional microplate reader: Varioskan Flash model, manufactured by Thermo scientific, imported 96-well plates, etc.

[0085] 5. Experimental method:

[0086] 1) When the U87 cells and A172 cells are cultured to the logarithmic growth phase, use 0.25% trypsin containing EDTA to digest the cells and collect them, and adjust the cell density in the cell suspension to the target value by cell counting.

[0087] 2) Add 100 μL of culture medium to each well of the 96-well plate, so that there are about 6000 cells in each well. The specific number of cells is determined according to factors such as cell size and proliferation rate. Set 3 replicates for each group, gently shake evenly, and then place the 96-well plate in a cell culture incubator at 37°C and 5% CO2 for culture.

[0088] 3) When the cells are in good adherent state, add the target drug to each well of the 96-well plate to the target concentration for drug stimulation, and then place the 96-well plate in an incubator at 37°C with 5% CO2 for 72 h.

[0089] 4) After reaching the target treatment time, add the pre-prepared CCK-8 solution melted at room temperature in the dark to each well of the 96-well plate, adding 10 μL of CCK-8 to every 100 μL of culture medium. Then place it in an incubator at 37°C with 5% CO2 in the dark for 0.5 - 1 h.

[0090] 5) Shake for 1 min in a multi-functional microplate reader, and use the multi-functional microplate reader to measure the absorbance value of each well at double absorption wavelengths of 450 nm and 620 nm. The results of the CCK-8 assay are expressed as the relative percentage of the absorbance value of the control group. When calculating the results, note to zero with the blank control well, record the experimental results and calculate the cell viability.

[0091] The results of the in vitro non-solid tumor cell inhibitory activity are shown in Table 1:

[0092] Table 1 Inhibitory Activity of Glioblastoma Cells in Vitro

[0093]

[0094]

[0095] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A curcuminol derivative containing a reactive Michael receptor, wherein the specific structural formula of the curcuminol derivative is as follows:

2. A method for preparing the derivatives S1, S5, S6, S9-S14 according to claim 1, characterized in that: Curcumol first reacts with mCPBA to form an epoxy intermediate. Then, under heating and alkaline conditions, the epoxy compound undergoes ring opening and elimination to produce a hydroxy compound II. Compound II reacts with a Dess-Martin oxidant and is oxidized to produce a derivative S1. Derivative S1 is oxidized under the action of sodium chlorite and hydrogen peroxide to produce a compound S2. Compound S2 undergoes a condensation reaction with various amines to produce derivatives described in S5, S6, and S9-S14:

3. A method for preparing the derivatives S18 and S19 according to claim 1, characterized in that: At room temperature, hydrated ruthenium trichloride was used as a catalyst, and curcumol was oxidized by sodium periodate to produce compound III. Compound III then underwent an aldol condensation reaction with paraformaldehyde to obtain derivative S18. Derivative S18 then underwent an addition reaction with dimethylamine to obtain S19.

4. Use of the curcumol derivative according to claim 1 in the preparation of a medicament for treating glioblastoma.

5. The use according to claim 4, characterized in that The active ingredient in the medicine is the curcuminol derivative according to claim 1.

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