Diphyllin ether 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 ether derivative as well as a preparation method and application thereof. The structural formula of the diphyllin ether derivative is shown in the specification, wherein R represents one of methyl, ethyl, allyl, isopropyl, tertiary butyl, cyclopropyl and benzyl. The diphyllin ether derivative is a diphyllin ether derivative with a non-glycoside structure, does not contain glycosidic bonds, double bonds and other structures which are easy to hydrolyze in vivo, has better metabolic stability than glycoside compounds, has higher tumor cell proliferation inhibition activity, and can be applied to preparation of medicines for preventing and treating colorectal adenocarcinoma, liver cancer and lung cancer.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical chemistry and pharmacology, and particularly relates to a cleistanthus root leaf ether derivative, a preparation method and application thereof. BACKGROUND

[0002] Cancer is a major public health problem that seriously endangers human health at present, and the exploration and research of plant-derived antitumor components is one of the core directions in this field. Cleistanthin, as a natural lignan extracted from the plant of Cleistanthus collmnsianus, is the key pharmacodynamic skeleton of its numerous active derivatives. This compound itself has biological activities such as antiviral activity, but as an antitumor lead compound, its direct activity is limited. Further research found that a series of glycoside derivatives (such as cleistanthin-A) obtained by structural modification of cleistanthin showed significantly enhanced antitumor activity, showing broad development prospects (Chinese Journal of Chemistry, 2007, 25, 679-682, Chemical Biology and Drug Design, 2015, 86, 691-696). Although the activity is significant, such glycoside derivatives have poor metabolic stability due to the complex synthesis route and the glycoside bond is easily hydrolyzed by enzymes in vivo. Therefore, pharmaceutical researchers designed and synthesized 4-C linked cleistanthin triazole derivatives to improve their stability. Unfortunately, although the metabolic stability of this new derivative is greatly improved, its antitumor activity is significantly reduced. (Chemical Biology & Drug Design, 2024, 104, e14635).

[0003] SUMMARY

[0004] Therefore, the purpose of the present application is to provide a cleistanthin ether derivative, a preparation method and application thereof. The cleistanthin ether derivative is a non-glycoside structure cleistanthin ether derivative, which does not contain glycoside bonds and double bonds that are easily hydrolyzed in vivo, has better metabolic stability than various glycoside derivatives of cleistanthin, and has strong tumor cell proliferation inhibition activity.

[0005] In a first aspect, the present application provides a cleistanthin ether derivative, which has the structure shown in formula (I):

[0006]

[0007] wherein R represents one of methyl, ethyl, allyl, isopropyl, tert-butyl, cyclopropyl and benzyl.

[0008] In some embodiments of the present application, the quercetin ether derivative has a structure as shown in any one of formulae 3a-3d:

[0009]

[0010] wherein,

[0011] When R is methyl, the quercetin ether derivative is a compound having a structure as shown in formula 3a;

[0012] When R is ethyl, the quercetin ether derivative is a compound having a structure as shown in formula 3b;

[0013] When R is allyl, the quercetin ether derivative is a compound having a structure as shown in formula 3c;

[0014] When R is isopropyl, the quercetin ether derivative is a compound having a structure as shown in formula 3d.

[0015] In a second aspect of the present application, a preparation method of a quercetin ether derivative is provided, comprising the following steps:

[0016] S1. obtaining quercetin epoxy ether 2 by reacting quercetin with epoxy bromopropane, potassium carbonate in N,N-dimethylformamide;

[0017] S2. obtaining quercetin ether derivative 3 by nucleophilic substitution reaction of quercetin epoxy ether 2 with aluminum triflate in alcohol ROH;

[0018] wherein, the reaction formula of the above preparation method is:

[0019]

[0020] wherein, R represents one of methyl, ethyl, allyl, isopropyl, tert-butyl, cyclopropyl and benzyl.

[0021] In some embodiments of the present application, the above preparation method comprises the following steps:

[0022] (1) dissolving quercetin in N,N-dimethylformamide, stirring at 25°C for 5 minutes, slowly adding epoxy bromopropane and potassium carbonate into the mixed solution to obtain a first reaction liquid; quenching the reaction by adding the first reaction liquid into deionized water, extracting with ethyl acetate, collecting the organic phase and sequentially washing with water, saturated brine, drying with anhydrous MgSO4, concentrating under reduced pressure, and then flash column chromatography to obtain a light yellow solid, i.e. quercetin epoxy ether 2;

[0023] (2) The reaction tube is added with shanhuhuigenin epoxy ether 2, aluminum triflate and alcohol ROH, and heated to react under nitrogen protection to obtain a second reaction solution. After the second reaction solution is cooled, it is concentrated under reduced pressure and then flash chromatography is performed to obtain white solid, i.e. shanhuhuigenin ether derivative 3;

[0024] In some embodiments of the present application, in the preparation method step S1, the molar ratio of shanhuhuigenin, epoxy bromopropane and potassium carbonate is 1:5:6; the reaction temperature is 40-70°C, and the reaction time is 1-3 hours.

[0025] In some embodiments of the present application, in the preparation method step S2, the molar ratio of shanhuhuigenin epoxy ether 2, alcohol ROH and aluminum triflate is 1:1.5:0.1; the reaction temperature is 40-70°C, and the reaction time is 2-4 hours.

[0026] In a third aspect of the present application, the shanhuhuigenin ether derivative is used for preparing a drug for treating cancer, and the cancer is one of liver cancer, colorectal adenocarcinoma and lung cancer.

[0027] Compared with the prior art, the shanhuhuigenin ether derivative with non-glycoside structure provided in the present application does not contain glycoside bonds which are easy to be hydrolyzed in vivo in the structure of the compound, and the metabolic stability is better than that of glycosides. Through in vitro tumor cell proliferation inhibition experiments, it is found that the compound has strong tumor cell proliferation inhibition activity, which is equivalent to or better than that of paclitaxel, a positive control drug, and can be applied to the preparation of drugs for preventing and treating liver cancer, colorectal adenocarcinoma and lung cancer. BRIEF DESCRIPTION OF DRAWINGS

[0028] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments or prior art description will be briefly introduced below.

[0029] Figure 1 NMR of 3a provided in Embodiment 2 of the present application 1 H spectrum;

[0030] Figure 2 NMR of 3a provided in Embodiment 2 of the present application 13 C spectrum;

[0031] Figure 3 NMR of 3b provided in Embodiment 3 of the present application 1 H spectrum;

[0032] Figure 4 NMR of 3b provided in Embodiment 3 of the present application 13 C spectrum;

[0033] Figure 5NMR of 3c provided for Example 4 of the invention 1 H spectrum;

[0034] Figure 6 NMR of 3c provided for Example 4 of the invention 13 C spectrum;

[0035] Figure 7 NMR of 3d provided for Example 5 of the invention 1 H spectrum;

[0036] Figure 8 NMR of 3d provided for Example 5 of the invention 13 C spectrum. DETAILED DESCRIPTION

[0037] The technical solutions in the embodiments of the present application will be described clearly and completely below. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all the other embodiments obtained by a person of ordinary skill in the art without creative work fall within the protection scope of the present application.

[0038] Example 1

[0039] Shanhuyexu (1.0 mmol), epibromohydrin (5.0 mmol), potassium carbonate (6.0 mmol) were dissolved in DMF, stirred at 60°C for 2 h, and the reaction was monitored by thin layer chromatography. After the reaction was completed, the water phase was extracted with ethyl acetate, and the organic phase was extracted with saturated sodium bicarbonate solution. The organic phase was dried with anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain yellow solid shanhuyexu epoxy ether 2 (0.85 mmol, 85%), R f = 0.5 (petroleum ether: ethyl acetate = 1:1). 1 HNMR (400 MHz, CDC13) 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, OCH20), 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). 13C NMR (101 MHz, CDC13) 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.

[0040] Example 2

[0041] Example 1, methanol (1.5 mmol) and aluminum trifluoromethanesulfonate (0.1 mmol) were added into a reaction tube under N2protection, stirred at 50 °C for 3 h, TLC monitored the reaction. After the reaction was completed, the residual methanol was removed by rotary evaporation, and the product was separated and purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain yellow solid 3a (0.72 mmol, 72%), Rf= 0.3 (petroleum ether: ethyl acetate = 1:1). f 1 H NMR (400 MHz, CDC13) δ 7.57 (s, 1H, ArH), 7.00 (s, 1H, ArH), 6.89 (d, J = 7.9 Hz, 1H, ArH), 6.76-6.67 (m, 2H, ArH), 6.00 (dd, J = 19.3, 1.5 Hz, 2H, OCH20), 5.40 (s, 2H, OCH2), 4.17 (ddt, J = 14.4, 9.4, 4.4 Hz, 3H, CH2CH), 4.00 (s, 3H, OCH3), 3.74 (s, 3H, OCH3), 3.63-3.55 (m, 2H, CH2), 3.39 (s, 3H, OCH3). 13 CNMR (101 MHz, CDC13) δ 168.63, 150.66, 149.21, 146.41, 145.53, 134.16, 129.69, 127.28, 125.91, 125.59, 122.55, 118.15, 109.66, 107.16, 105.16, 100.20, 99.41, 72.69, 72.11, 68.52, 65.37, 58.38, 55.07, 54.80, 28.67. HRMS (ESI): m / z calcd for C 25 H 25 O9: 469.1504; found: 469.1499 [M+H] + .

[0042] NMR of white solid 3a 1 ​H spectrum as shown Figure 1 As shown; nuclear magnetic resonance 13 C spectrum as shown Figure 2 As shown.

[0043] Example 3

[0044] Under N2 protection, 1.0 mmol of kaempferol epoxy ether 2, 1.5 mmol of ethanol, and 0.1 mmol of aluminum trifluoromethanesulfonate were added to a reaction tube and stirred at 50 °C for 3 h. The reaction was monitored by thin-layer chromatography. After the reaction was complete, the remaining ethanol was removed by rotary evaporation, and the product was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to give a white solid 3b (0.82 mmol, 82%). f =0.3 (petroleum ether: ethyl acetate = 1:1). 1 H NMR (400MHz, CDCl3) δ7.65 (s, 1H, ArH), 7.07 (s, 1H, ArH), 6.96 (d, J = 7.8Hz, 1H, ArH), 6.84 -6.76(m,2H,ArH),6.08(dd,J=19.3,1.4Hz,2H,OCH2O),5.49(d,J=5.7Hz,2H,OCH2),4.25( tdd,J=16.2,9.3,5.5Hz,3H,CH2CH),4.07(s,3H,OCH3),3.81(s,3H,OCH3),3.70(td,J=9. 2,4.8Hz,2H,CH2),3.61(dtd,J=9.2,7.0,6.1,2.7Hz,2H,CH2),1.24(d,J=7.2Hz,3H,CH3). 13 C NMR (101MHz, CDCl3) δ169.69,151.70,150.27,147.50,146.64,135.17,130.74,128.35,126.90,126.65,123.62,119. 22,110.73,108.22,106.20,101.25,100.53,73.83,71.01,69.67,67.15,66.46,56.13,55.86,15.16.HRMS(ESI):m / z calcd for C 26 H 27 O9:483.1659; found:483.1655[M+H] + .

[0045] Nuclear magnetic resonance of white solid 3b 1 H spectrum as shown Figure 3 As shown; nuclear magnetic resonance 13 C spectrum as shownFigure 4

[0046] Example 4

[0047] Under N2protection, the reaction tube was charged with the maytea oroxylin epoxy ether 2 (1.0 mmol), propargyl alcohol (1.5 mmol) and aluminum trifluoromethanesulfonate (0.1 mmol), stirred at 50 °C for 3 h, TLC monitored the reaction. After the reaction was completed, the residual propargyl alcohol was removed by rotary evaporation, the product was isolated and purified by column chromatography (petroleum ether: ethyl acetate = 1 : 1) to give white solid 3c (0.73 mmol, 73%), Rf= 0.3 (petroleum ether: ethyl acetate = 2: 1). f 1 H NMR (400 MHz, CDC13) δ 7.64 (s, 1H, ArH), 7.07 (s, 1H, ArH), 6.95 (d, J = 7.8 Hz, 1H, ArH), 6.83-6.76 (m, 2H, ArH), 6.07 (dd, J = 19.5, 1.4 Hz, 2H, OCH20), 5.92 (ddd, J = 16.4, 10.8, 5.4 Hz, 1H, CH), 5.47 (s, 2H, OCH2), 5.35-5.17 (m, 2H, CH2), 4.32-4.19 (m, 3H, CH2CH), 4.09 (dt, J = 5.7, 1.4 Hz, 2H, CH2), 4.06 (s, 3H, OCH3), 3.81 (s, 3H, OCH3), 3.71 (qd, J = 9.5, 5.0 Hz, 2H, CH2). 13 C NMR (101 MHz, CDC13) δ 169.72, 151.69, 150.26, 147.48, 146.60, 135.15, 134.10, 130.71, 128.34, 126.88, 126.63, 123.61, 119.18, 117.81, 110.72, 108.20, 106.19, 101.25, 100.51, 73.78, 72.55, 70.66, 69.68, 66.48, 56.14, 55.85. HRMS (ESI): m / z calcd for C 27 H 27 O 9: 495.1656; found: 495.1655 [M+H] + .

[0048] The NMR spectrum of white solid 3c 1 H spectrum is shown as Figure 5 ; the NMR spectrum of C is shown as 13 Figure 6 . ​​​

[0049] Example 5

[0050] Under N2protection, the reaction tube was charged with the ginkgolid epoxy ether 2 (1.0 mmol), isopropanol (1.5 mmol) and aluminum trifluoromethanesulfonate (0.1 mmol), stirred at 50 °C for 3 h, TLC monitored the reaction. After the reaction was completed, the residual isopropanol was removed by rotary evaporation, the product was isolated and purified by column chromatography (petroleum ether: ethyl acetate = 1 : 1) to give white solid 3d (0.73 mmol, 73%), Rf= 0.3 (petroleum ether: ethyl acetate = 1 : 1). f 1 H NMR (400 MHz, CDC13) δ 7.66 (s, 1H, ArH), 7.07 (s, 1H, ArH), 6.96 (d, J = 7.9 Hz, 1H, ArH), 6.87 - 6.77 (m, 2H, ArH), 6.08 (dd, J = 19.9, 1.5 Hz, 2H, OCH20), 5.49 (s, 2H, OCH2), 4.30 - 4.15 (m, 3H, CH2CH), 4.07 (s, 3H, OCH3), 3.81 (s, 3H, OCH3), 3.70 (d, J = 6.5 Hz, 1H, CH), 3.65 (ddd, J = 15.0, 8.5, 3.5 Hz, 2H, CH2), 1.21 (s, 3H, CH3), 1.20 (s, 3H, CH3). 13 C NMR (101 MHz, CDC13) δ 169.75, 151.69, 150.27, 147.50, 146.69, 135.13, 130.74, 128.36, 126.82, 126.66, 123.62, 119.21, 110.73, 108.22, 106.20, 101.26, 100.57, 73.86, 72.59, 69.81, 68.58, 66.52, 56.14, 55.86, 29.73, 22.08. HRMS (ESI): m / z calcd for C 27 H 29 O9: 497.1816; found: 497.1812 [M+H] + .

[0051] The NMR spectrum of white solid 3d 1 H as shown; the NMR spectrum of Figure 7 C as shown. 13 Figure 8

[0052] ​​​In order to better understand the essence of the present application, the following pharmacological experimental results of the growth inhibition of three tumor cell lines by the shanzhistane ether derivatives provided by the present application are used to illustrate the new use of the shanzhistane ether derivatives in the field of anti-tumor drug research. The pharmacological examples give some activity data of representative compounds. It must be pointed out that the pharmacological examples of the present application are used to illustrate the present application and are not a limitation of the present application. Simple improvements of the present application according to the essence of the present application are within the scope of the present application.

[0053] Pharmaceutical experiment example 1

[0054] Cytotoxic activity test of compounds 3a-3d and paclitaxel on human hepatoma cells (HepG2), human colorectal adenocarcinoma cells (HCT-15), and human lung cancer cells (A549)

[0055] Human hepatoma cells (HepG2), human colorectal adenocarcinoma cells (HCT-15), 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.

[0056] Cells in the logarithmic growth phase were inoculated in a 96-well plate at a density of 5×10 3 cells per well, and cultured for 24 hours to allow the cells to adhere fully. The test compound was dissolved in DMSO to prepare a 1×10 -2 mol / L stock solution, which was diluted with complete medium to the corresponding concentration to obtain a test compound solution of different concentrations. After removing the original culture medium, the culture medium containing different concentrations of compound 3a was added, with 4 parallel holes set for each concentration, and the culture was continued for 68 hours. After the culture was completed, tetramethyl azo salt (MTT) solution was added to each well, and the culture was continued for 4 hours. The culture medium was discarded, 150 μL of dimethyl sulfoxide was added to each well, and the mixture was shaken for 10 minutes. 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.

[0057] Table 1 Cytotoxic activity test results of compounds 3a-3d and paclitaxel

[0058]

[0059] According to Table 1, the shanhoye ether derivative provided by the application has important biological activity, in-vitro cell toxicity activity test on three kinds of tumor cells, human hepatoma (HepG2), human lung cancer cells (A549), human colorectal adenocarcinoma cells (HCT-15) and the like, shows that the shanhoye ether derivative provided by the application has inhibitory effect on tumor cell growth, and is possible to develop into a new tumor prevention and treatment drug. From the above pharmacological examples, it can be seen that these compounds show strong cell toxicity activity on the three kinds of tumor cells, and the cell toxicity activity of most of the compounds is close to the positive control paclitaxel, and has the potential to develop into an anti-tumor drug.

[0060] While some embodiments of the general inventive concept have been shown and described, it is to be understood that changes can be made in these embodiments without departing from the principles and spirit of the general inventive concept, the scope of which is defined in the claims and their equivalents.

Claims

1. A safflower leaf extract ether derivative, characterized in that, The structural formula of the kaempferol leaf extract derivative is shown below: Wherein, R represents one of methyl, ethyl, allyl, isopropyl, tert-butyl, cyclopropyl, and benzyl.

2. The physalis leaf extract ether derivative according to claim 1, characterized in that, The kaempferol ether derivative has a structure as shown in any one of formulas 3a-3d:

3. A method for preparing the senna leaf extract ether derivative 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 2. S2. The sorbitol epoxy ether 2 and aluminum trifluoromethanesulfonate underwent a nucleophilic substitution reaction in ROH to obtain sorbitol ether derivative 3. The reaction formula for the above preparation method is as follows: Wherein, R represents one of methyl, ethyl, allyl, isopropyl, tert-butyl, cyclopropyl, and benzyl.

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 senna leaf extract epoxy ether 2, ROH and aluminum trifluoromethanesulfonate is 1:1.5:0.1; the reaction temperature is 40℃-70℃ and the reaction time is 2-4 hours.

6. The use of the senna leaf extract derivative as described in claim 1 in the preparation of a medicament for treating cancer, wherein the cancer is one of liver cancer, colorectal adenocarcinoma, and lung cancer.

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