Trimethoxy phenyl derivative as well as preparation method and application thereof

Abstract

The invention relates to a trimethoxy phenyl derivative as well as a preparation method, a pharmaceutical composition and application thereof. Specifically disclosed is a compound represented by formula I or a pharmaceutically acceptable salt thereof. The compound provided by the invention has good tumor inhibition activity.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a trimethoxyphenyl derivative, its preparation method, and its application. Background Technology

[0002] Malignant tumors, as a major threat to human health, have seen a continuous upward trend in both global incidence and mortality rates.

[0003] Some natural products containing trimethoxy groups have significant antitumor activity, such as podophyllotoxin and colchicine.

[0004]

[0005] Clinically, the treatment of malignant tumors mainly relies on chemotherapy, surgery, radiotherapy, hormone therapy, and immunotherapy. Among these, chemotherapy, as one of the most commonly used treatments, uses anticancer drugs to destroy or kill cancer cells, thereby inhibiting tumor growth. However, while chemotherapy drugs exert their anti-tumor effects, they are also accompanied by widespread toxic side effects and drug resistance. These drugs lack selectivity for both tumor cells and normal cells, leading to damage to normal cells while killing or inhibiting tumor cells, thus affecting the function of the heart, liver, kidneys, and nervous system. Therefore, the development of low-toxicity and highly effective anticancer drugs is particularly important in cancer treatment. Summary of the Invention

[0006] Objectives of the invention: The first objective of this invention is to provide a trimethoxyphenyl derivative; the second objective of this invention is to provide a method for preparing the trimethoxyphenyl derivative; and the third objective of this invention is to provide applications of the trimethoxyphenyl derivative.

[0007] Technical solution: The trimethoxyphenyl derivative with antitumor activity described in this invention is a compound with the general structural formula as shown in formula (I) or its salt, stereoisomer, or hydrate:

[0008]

[0009] Among them, Linker is independently selected from C1-C4 alkane chains and C1-C4 olefin chains;

[0010] R 1 for Trifluoromethyl, halogen, alkyl.

[0011] The compound is a compound having any of the following general structural formulas or a salt, stereoisomer, or hydrate thereof:

[0012]

[0013] The preparation method includes the following steps: compound 1 and compound 2 are subjected to an amidation reaction to obtain compound I:

[0014]

[0015] Where n is 1 to 5.

[0016] The condensing agent for the amidation reaction described in the above steps is 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (HATU), benzotriazole-N,N,N′,N′-tetramethylurea hexafluorophosphate (HBTU), 1-butylphosphonic anhydride (T4P), O-benzotriazole-N,N,N′N′-tetramethylurea tetrafluoroborate (TBTU), tetramethylchlorourea hexafluorophosphate (TCFH), or carbonyl diimidazole (CDI). The reaction solvent is (1-cyano-2-ethoxy-2-oxoethyleneaminooxy)dimethylamino-morpholine-carbomony hexafluorophosphate (COMU); preferably TCFH; the reaction solvent is dimethyl sulfoxide, N,N-dimethylformamide, dichloromethane, acetonitrile, anhydrous ethanol, preferably acetonitrile; the reaction base is potassium carbonate, cesium carbonate, triethylamine, N,N-diisopropylethylamine (DIPEA), N-methylimidazole (NMI), preferably N-methylimidazole (NMI).

[0017] The application of the trimethoxyphenyl derivative described in this invention in the treatment of tumors.

[0018] The tumors are breast cancer and kidney cancer.

[0019] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: the trimethoxyphenyl derivative of the present invention has good anti-tumor activity. Detailed Implementation

[0020] The technical solution of the present invention will be further described below with reference to the embodiments.

[0021] Example 1

[0022] The trimethoxyphenyl derivative of this invention, chemically named N-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)-3,4,5-trimethoxybenzoyl, has the following structural formula:

[0023]

[0024] The preparation method is as follows: 176 mg (0.83 mmol) of 3,4,5-trimethoxybenzoic acid, 317 mg (1.13 mmol) of TCFH, 262 μL (2.40 mmol) of NMI, 200 mg (0.94 mmol) of 3-chloro-4-(pyridin-2-ylmethoxy)aniline, and 3 mL of acetonitrile were added sequentially to a reaction flask. The mixture was stirred at 25℃~35℃ for 2 h. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 80:1) to obtain 158 mg of the target product, with a yield of 44.4%.

[0025] The target compound is a white solid; 1 H NMR(400MHz, DMSO-d6)δ 10.30 (s, 1H), 8.87 (s, 2H), 8.59 (d, J = 4.2Hz, 1H), 7.97 (d, J = 2.5Hz, 1H), 7.87 (td, J = 7.7, 1.8Hz, 1H), 7.69 (dd, J = 9.0, 2.6Hz , 1H), 7.36 (dd, JJ=7.6, 4.9Hz, 1H), 7.32 (s, 1H), 7.25 (d, J=9.2Hz, 1H), 5.27 (s, 2H), 3.87 (s, 3H), 3.83 (s, 8H), 3.72 (s, 3H). 13 C NMR(101MHz, DMSO-d6)δ 165.16, 156.82, 153.08, 150.07, 149.59, 140.80, 137.56, 136.48, 133.72, 130.12, 123.5 1, 123.21, 122.76, 121.89, 121.33, 120.91, 114.75, 105.81, 71.63, 60.58, 56.62, 35.47.

[0026] Example 2

[0027] The trimethoxyphenyl derivative of this invention has the chemical name (E)-N-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)-3-(3,4,5-trimethoxyphenyl)acrylamide, and its structural formula is as follows:

[0028]

[0029] The preparation method is as follows: 198 mg (0.83 mmol) of (E)-3-(3,4,5-trimethoxyphenyl)acrylic acid, 317 mg (1.13 mmol) of TCFH, 262 μL (2.40 mmol) of NMI, 200 mg (0.94 mmol) of 3-chloro-4-(pyridin-2-ylmethoxy)aniline, and 3 mL of acetonitrile were added sequentially to a reaction flask. The mixture was stirred at 25℃~35℃ for 2 h. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 80:1) to obtain 180 mg of the target product, with a yield of 47.7%.

[0030] The target compound is a white solid; 1 H NMR(400MHz,DMSO-d6)δ 9.35 (s, 1H), 7.70 (dd, J=4.7, 1.6Hz, 1H), 7.10 (s, 0H), 7.02-6.94 (m, 1H), 6.70-6.65 (m, 1H), 6.64-6.59 (mm, 1H), 6. 50-6.43 (m, 1H), 6.33 (d, J=9.0Hz, 1H), 6.08 (s, 2H), 5.83 (d, J=15.6Hz, 1H), 4.37 (s, 2H), 2.95 (s, 6H), 2.82 (s, 3H). 13 C NMR (101MHz, DMSO-d6) δ163.94, 156.84, 153.58, 149.80, 149.56, 140.89, 139.46, 137.52, 134.06 , 130.70, 123.47, 121.83, 121.74, 121.71, 121.20, 119.36, 115.07, 105.64, 71.66, 60.57, 56.35.

[0031] Example 3

[0032] The trimethoxyphenyl derivative of this invention, chemically named N-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)-2-(3,4,5-trimethoxyphenyl)acetamide, has the following structural formula:

[0033]

[0034] The preparation method is as follows: 188 mg (0.83 mmol) of 2-(3,4,5-trimethoxyphenyl)acetic acid, 317 mg (1.13 mmol) of TCFH, 262 μL (2.40 mmol) of NMI, 200 mg (0.94 mmol) of 3-chloro-4-(pyridin-2-ylmethoxy)aniline, and 3 mL of acetonitrile were added sequentially to a reaction flask. The mixture was stirred at 25℃~35℃ for 2 h. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 80:1) to obtain 117 mg of the target product, with a yield of 31.8%.

[0035] The target compound is a white solid; 1 H NMR(400MHz, DMSO-d6)δ 10.13 (s, 1H), 8.58 (ddd, J=5.0, 1.8, 1.0Hz, 1H), 7.89-7.81 (m, 2H), 7.54 (d, J=7.8Hz, 1H), 7.42 (dd, J=8.9, 2.6Hz, 1H), 7 .34 (ddd, J=7.6, 4.9, 1.2Hz, 1H), 7.18 (d, J=9.0Hz, 1H), 6.64 (s, 2H), 5.23 (s, 2H), 3.77 (s, 6H), 3.64 (s, 3H), 3.55 (s, 2H). 13 C NMR(101MHz, DMSO)δ 169.38, 156.82, 153.16, 149.73, 149.56, 137.51, 136.78, 133.87, 131.80, 123.4 7, 121.82, 121.64, 121.29, 119.40, 114.99, 106.99, 71.64, 60.43, 56.30, 43.92.

[0036] Example 4

[0037] The trimethoxyphenyl derivative of this invention, chemically named 3,4,5-trimethoxy-N-(3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)benzamide, has the following structural formula:

[0038]

[0039] The preparation method is as follows: 176 mg (0.83 mmol) of 3,4,5-trimethoxybenzoic acid, 317 mg (1.13 mmol) of TCFH, 262 μL (2.40 mmol) of NMI, 3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)aniline, and 3 mL of acetonitrile were added sequentially to a reaction flask. The mixture was stirred at 25℃~35℃ for 2 h. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 80:1) to obtain 158 mg of the target product, with a yield of 43.7%.

[0040] The target compound is a white solid; 1 H NMR(400MHz, DMSO-d6)δ 10.54 (s, 1H), 8.28 (t, J = 2.0Hz, 1H), 8.21 (d, J = 1.4Hz, 1H), 8.09 (t, J = 1.8Hz, 1H), 7.73 (t, J = 2 .0Hz, 1H), 7.49 (t, J = 1.3Hz, 1H), 7.33 (s, 2H), 3.89 (s, 6H), 3.75 (s, 3H), 2.18 (d, J = 1.0Hz, 3H). 13 C NMR(101MHz, DMSO-d6)δ 165.84, 153.20, 141.70, 141.29, 139.41, 138.42, 135.44, 131.46, 131.14, 129.55, 12 5.47, 122.76, 115.55, 114.93, 114.67, 112.14, 112.11, 105.95, 60.64, 56.65, 14.01.

[0041] Example 5

[0042] The trimethoxyphenyl derivative of this invention has the chemical name (E)-N-(3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-(3,4,5-trimethoxyphenyl)acrylamide, and its structural formula is as follows:

[0043]

[0044] The preparation method is as follows: 198 mg (0.83 mmol) of (E)-3-(3,4,5-trimethoxyphenyl)acrylic acid, 317 mg (1.13 mmol) of TCFH, 262 μL (2.40 mmol) of NMI, 3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)aniline, and 3 mL of acetonitrile were added sequentially to a reaction flask. The mixture was stirred at 25℃~35℃ for 2 h. The reaction solution was concentrated and purified by column chromatography (dichloromethane:methanol = 80:1) to obtain 67 mg of the target product, with a yield of 17.5%.

[0045] The target compound is a white solid; 1 H NMR(400MHz, DMSO-d6)δ 10.67 (s, 1H), 8.18 (d, J = 1.4Hz, 1H), 8.10 (t, J = 2.0Hz, 1H), 8.02 (d, J = 2.1Hz, 1H), 7.67 (d, J = 2.3Hz, 1H), 7.59 (d , J=15.6Hz, 1H), 7.48-7.43 (m, 1H), 6.98 (s, 2H), 6.74 (d, J=15.7Hz, 1H), 3.84 (s, 6H), 3.70 (s, 3H), 2.18 (s, 3H). 13 C NMR(101MHz, DMSO-d6)δ 164.82, 153.63, 142.04, 141.89, 140.07, 139.80, 139.41, 138.61, 135.41, 130.43, 125.42, 1 22.71, 121.25, 114.71, 114.37, 113.74, 111.89, 106.13, 105.90, 60.61, 56.54, 56.43, 14.01.

[0046] Example 6

[0047] The trimethoxyphenyl derivative of this invention has the chemical name (E)-N-(3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-(3,4,5-trimethoxyphenyl)acrylamide, and its structural formula is as follows:

[0048]

[0049] The preparation method is as follows: 188 mg (0.83 mmol) of 2-(3,4,5-trimethoxyphenyl)acetic acid, 317 mg (1.13 mmol) of TCFH, 262 μL (2.40 mmol) of NMI, 200 mg (0.94 mmol) of 3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)aniline, and 3 mL of acetonitrile were added sequentially to a reaction flask. The mixture was stirred at 25℃~35℃ for 2 h. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 80:1) to obtain 80 mg of the target product, with a yield of 21.4%.

[0050] The target compound is a white solid; 1H NMR(400MHz, DMSO-d6)δ 10.66 (s, 1H), 8.13 (d, J = 1.4Hz, 1H), 8.02 (t, J = 2.0Hz, 1H), 7.91 (d, J = 1.9Hz, 1H), 7.63 (d, J = 2 .0Hz, 1H), 7.41 (t, J=1.3Hz, 1H), 6.63 (s, 2H), 3.74 (s, 6H), 3.60 (s, 5H), 2.13 (d, J=1.0Hz, 3H). 13 C NMR(101MHz, DMSO-d6)δ 170.42, 153.19, 141.77, 139.37, 138.51, 136.87, 135.43, 131.57, 131.28, 125.40, 1 22.68, 114.67, 114.23, 113.57, 111.76, 111.73, 107.17, 60.43, 56.34, 44.01, 14.00.

[0051] Bioactivity evaluation

[0052] MTT assay for antitumor activity

[0053] Cell culture: Human cancer cell lines MDA-MB-231 and A498 were purchased from the National Biomedical Laboratory in Beijing and cultured in DMEM (KGM12800-500) or MEM (KGM41500-500) medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin, at 37°C in a Thermo Fisher Scientific (BB150) incubator containing 5% CO2. When the cell confluence reached 70%-80%, 0.25% trypsin was added for digestion, resuspending, and culturing. Cells in the logarithmic growth phase and in good growth condition were selected for further study.

[0054] Methyl thiazolyl tetrazolium (MTT) was used to determine cell viability. Hemocytometer counting was used for cell counting, and cell viability was greater than 95% in all experiments. MDA-MB231 and A498 cells were cultured at 1 × 10⁻⁶ cells / mL. 4Cells were seeded in 96-well plates. 100 μL of medium (containing 1% FBS) was added to each well to dissolve different concentrations of the drug (0-50 μM), and the cells were incubated for 24 h. After centrifugation (5 min, 2000 rpm), the supernatant was discarded, and 10 μL of LTT (5 mg / mL) solution was added to each well. The cells were incubated at 37°C for 4 h, centrifuged again, and the supernatant was discarded. 100 μL of DMSO was added to each well, and the cells were shaken for 10 min to fully dissolve the formazan crystals. The absorbance was measured at 570 nm using a microplate reader (BioTek, USA). Cytotoxicity was assessed compared to the control group (DMSO). The concentration at which the drug induced 50% cell growth inhibition (IC50) was determined using a curve fitting algorithm in GraphPad Prism 9 (GraphPad software, LaJolla, CA, USA) via nonlinear regression. 50 ).

[0055] To investigate the effects of the compounds on tumor cell activity, cytotoxicity was detected using the MTT assay. Table 1 shows the effects of the compounds on the activity of different tumor cell lines, and the half-maximal inhibitory concentration (IC50) of the drugs was calculated for each cell line. 50 (48h).

[0056] Table 1. Inhibitory activity of compounds against different tumor cells (IC50) 50 )

[0057] Example MDA-MB-231(μM) A498 (μM) Example 1 20.31±0.60 11.19±0.53 Example 2 7.23±0.21 6.19±0.26 Example 3 / 5.83±0.24 Example 4 38.79±3.94 13.29±1.13 Example 5 19.69±0.10 27.15±3.69 Example 6 / 12.09±0.81 Podophyllotoxin 69.44±5.47 68.52±10.43

[0058] " / " indicates that it was not tested.

[0059] The results in Table 1 show that the compound has a significant inhibitory effect on MDA-MB-231 and A498 tumor cells, and has great research value.

Claims

1. A compound of formula (I), or a salt thereof, or a hydrate thereof: in, Linker is independently selected from C1-C4 alkane chains and C1-C4 olefin chains; n is 1 to 5; R 1 for Trifluoromethyl, halogen, alkyl.

2. The trimethoxyphenyl derivative with antitumor activity according to claim 1, characterized in that: It is a compound having any of the following general structural formulas or its salt, stereoisomer, or hydrate:

3. A method for preparing the trimethoxyphenyl derivative of claim 1, characterized in that, The following steps are included: Compound 1 and Compound 2 are amidated to obtain Compound I: Where n is 1 to 5.

4. A pharmaceutical composition, characterized in that, It includes the compounds as described in claims 1 to 2 and at least one pharmaceutically acceptable excipient.

5. The use of a substance in the preparation of a medicament for treating diseases, characterized in that, The substance is a compound as described in claims 1-2, and the disease is cancer; preferably breast cancer and kidney cancer.

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