Derrone phenylchalcone amide derivatives, their preparation and medical use

Abstract

The application discloses a pterostilbene paeonol chalcone amide derivative shown in a general formula (I) or a pharmaceutically acceptable salt thereof, the compound has a good inhibiting effect on human lung cancer cell A549, human hepatoma cell HepG2, colon cancer cell HCT116 and human osteosarcoma cell U-2OS tumor cell, and shows a certain inhibiting effect on HDAC1, 2, 3, 6 and 8, and has the potential of being used as an HDAC inhibitor type antitumor drug. The method for preparing the compound has the characteristics of easy availability of raw materials, simple operation and high yield.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of drug synthesis, in particular to a pterostilbene paeonol chalcone amide derivative, a preparation method and medical application thereof, the compound has excellent anti-tumor activity and can be used for preparing an anti-tumor drug. BACKGROUND

[0002] Pterostilbene is a stilbene compound, which is a homolog of resveratrol and mainly comes from blueberries and grapes. Existing researches have found that pterostilbene has multiple effects such as anti-cancer, anti-oxidation and anti-inflammation, and has small toxic and side effects. Researches have found that a large dose of pterostilbene (3000 mg / kg / d) used in animals also has no obvious toxic and side effects. Pterostilbene is a natural dimethylated analog of resveratrol, has a significant inhibitory effect on the proliferation of various tumor cells, and can inhibit the proliferation of various types of cancer, including lung cancer, breast cancer, prostate cancer, gastric cancer, colon cancer, pancreatic cancer, thyroid cancer, ovarian cancer and cervical cancer. The anti-tumor mechanism includes inducing tumor cell apoptosis, causing cell cycle arrest and blocking tumor cell growth and proliferation signal transduction. Researches have found that pterostilbene can regulate HDAC I, II and IV, and can also inhibit tumors by regulating SIRT. Pterostilbene is expected to develop into a new generation of anti-tumor drugs and gradually attract people's attention. Therefore, it has a good application prospect to develop pterostilbene HDAC inhibitors as chemical prophylactic drugs or cancer treatments.

[0003] However, due to the low structural stability, weak activity intensity, low bioavailability and unclear target of pterostilbene, the clinical application and medical value thereof are seriously limited.

[0004] Therefore, it is necessary to modify and transform the chemical structure of pterostilbene to overcome its limitations and improve the anti-cancer effect. SUMMARY

[0005] In view of the deficiencies of the prior art, the present application introduces an aldehyde group at position 4 of pterostilbene, and then combines the aldehyde group with an active ingredient paeonol to synthesize pterostilbene paeonol chalcone amide compounds, and studies the HDAC activity of the compounds. A series of new anti-tumor pterostilbene analogs which are safer and more effective are designed and synthesized, and have very important clinical application prospects and practical value.

[0006] In a first aspect, the present application provides a pterostilbene paeonol chalcone amide derivative or a pharmaceutically acceptable salt thereof as shown in the general formula (I):

[0007]

[0008] In the formula, R is selected from any one of a nitrogen-containing group, a C1-C6 alkylamine group and a C1-C6 alkanolamine group.

[0009] According to the specific embodiment of the present application, the nitrogen-containing group is a nitrogen-containing heterocyclic group, preferably any one of piperidyl, morpholinyl, piperazinyl, N-methylpiperazinyl, N-ethylpiperidyl, N-benzylpiperazinyl, N-hydroxyethylpiperazinyl, tetrahydropyrrolyl. The pharmaceutically acceptable salt includes, but is not limited to, sodium salt, potassium salt, calcium salt, magnesium salt, amino acid salt, sulfate salt, phosphate salt, maleate salt, fumarate salt, citrate salt, methanesulfonate salt, p-toluenesulfonate salt or tartrate salt.

[0010] In a first aspect of the present application, a preparation method of the aforementioned pterostilbene phloroglucinol chalcone amide derivative is provided, comprising the following steps:

[0011] S1, under alkaline conditions, a nucleophilic substitution reaction is performed on pterostilbene, a compound shown in formula (1) and a solvent to obtain a compound shown in formula (2), and the reaction formula is as follows:

[0012] wherein X is halogen;

[0013] S2, a Vilsmeier-Haack reaction is performed on the compound shown in formula (2), a formylating reagent and an acidic chloride to obtain a compound shown in formula (3), and the reaction formula is as follows:

[0014]

[0015] S3, under alkaline conditions, a hydrolysis reaction is performed on the compound shown in formula (3) and a solvent to obtain a compound shown in formula (4), and the reaction formula is as follows:

[0016]

[0017] S4, a nucleophilic substitution reaction is performed on the compound shown in formula (4), a compound shown in formula (5), a condensing agent and a solvent to obtain a compound shown in formula (6), and the reaction formula is as follows:

[0018]

[0019] The compound shown in formula (5) is selected from any one of a nitrogen-containing compound, a C1-C6 alkylamine compound and a C1-C6 alkanolamine compound; the definition of R is as described above;

[0020] S5, under alkaline conditions, a hydroxy aldehyde condensation reaction is performed on the compound shown in formula (6), phloroglucinol and a solvent to obtain a compound shown in formula (I), and the reaction formula is as follows:

[0021]

[0022] According to a specific embodiment of the present invention, in step S1, the molar ratio of pterostilbene to the compound represented by formula (1) is 1:1-3; preferably, the base includes potassium carbonate, and the solvent includes acetonitrile; preferably, the reaction temperature is 60-90° C., and the reaction time is 12-24 h.

[0023] According to a specific embodiment of the present invention, in step S2, the mass volume ratio of the compound represented by formula (2), the formylating agent and the acidic chloride is 1:1 to 3:1 to 3; preferably, the formylating agent is N,N-dimethylformamide; preferably, the reaction temperature is room temperature, and the reaction time is 2 to 4 hours.

[0024] According to a specific embodiment of the present invention, in step S3, the base includes sodium hydroxide solution, and the solvent includes ethanol; preferably, the reaction temperature is 40-60° C., and the reaction time is 1-4 hours.

[0025] According to a specific embodiment of the present invention, in step S4, the molar ratio of the compound represented by formula (4) to the compound represented by formula (5) is 1:1-3; preferably, the condensing agent includes HATU and DIPEA, and the solvent includes N,N-dimethylformamide; preferably, the reaction temperature is room temperature, and the reaction time is 1-3 hours.

[0026] According to a specific embodiment of the present invention, in step S5, the molar ratio of the compound represented by formula (6) to paeonol is 1:1-3; preferably, the base includes pyrrolidine, and the solvent includes ethanol; preferably, the reaction temperature is 30-60°C, and the reaction time is 12-24h.

[0027] The third aspect of the present invention provides a pharmaceutical composition, which is composed of a therapeutically effective amount of the aforementioned pterostilbene paeonol chalcone amide derivative or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier; preferably, the dosage form of the pharmaceutical composition is any one of ordinary tablets or capsules, sustained-release tablets or capsules, controlled-release tablets or capsules, oral solution, and injection.

[0028] The fourth aspect of the present invention provides the use of the aforementioned pterostilbene paeonol chalcone amide derivatives or pharmaceutically acceptable salts thereof in the preparation of anti-tumor drugs; preferably, the tumor is any one of lung cancer, breast cancer, prostate cancer, gastric cancer, colon cancer, pancreatic cancer, thyroid cancer, ovarian cancer, liver cancer, human osteosarcoma and cervical cancer; preferably, the drug is a histone deacetylase inhibitor.

[0029] Beneficial effects of the present invention:

[0030] The application designs and synthesizes a series of new-type structural pterostilbene paeonol chalcone amide compounds. The phenolic hydroxyl group in the structure of pterostilbene is easy to be oxidized. In view of this, a carboxyl group is introduced on the phenolic hydroxyl group, and then an amide is formed with different organic amines, so as to increase the water solubility. Then an aldehyde group is introduced at the 4-position, and then a chalcone compound is formed with paeonol having anti-tumor activity, so as to synergistically enhance the anti-tumor activity. The compound has good inhibitory effect on human lung cancer cell A549, human liver cancer cell HepG2, colon cancer cell HCT116 and human osteosarcoma cell U-2OS tumor cells, and shows certain inhibitory effect on HDAC1, 2, 3, 6 and 8, and has the potential as an HDAC inhibitor type anti-tumor drug. The method for preparing the compounds has the characteristics of easy-to-obtain raw materials, simple operation and high yield. DETAILED DESCRIPTION

[0031] In order to make the purpose, technical scheme and advantages of the embodiments of the application clearer, the technical scheme in the embodiments of the application will be clearly and completely described below with reference to the drawings in the embodiments of the application. Obviously, the described embodiments are some embodiments of the application, rather than all the embodiments. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative labor are within the protection scope of the application.

[0032] Example 1: Preparation of (E)-3-(2,4-dimethoxy-6-((E)-4-(2-oxo-2-(pyrrolidin-1-yl)ethoxy)styryl)phenyl)-1-(2-hydroxy-5-methoxyphenyl)prop-2-en-1-one (C-1)

[0033]

[0034] a. Pterostilbene (10.00 g, 39.02 mmol) was added to a round-bottom flask, then 150 mL of acetonitrile was added to the reaction bottle, and then anhydrous potassium carbonate (5.39 g, 39.02 mmol), ethyl bromoacetate (9.77 g, 58.52 mmol) were added. After stirring at 80°C for 20h, the reaction was basically completed by TLC monitoring. The solvent was recovered to dryness under reduced pressure, then 150 mL of ethyl acetate was added, washed with water (60 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a white product, which was intermediate 1. 1HNMR (400 MHz, CDC13) δ 7.44 (d, J = 8.6 Hz, 2H), 7.03 (d, J = 16.2 Hz, 1H), 6.94 - 6.88 (m, 3H), 6.67 - 6.62 (m, 2H), 6.38 (s, 1H), 4.63 (s, 2H), 4.28 (q, J = 7.1 Hz, 2H), 3.82 (s, 6H), 1.30 (t, J = 7.1 Hz, 3H). 13 C NMR (100 MHz, CDC13) δ 190.6, 168.8, 165.0, 164.5, 157.7, 142.9, 131.78, 131.0, 128.4, 126.1, 116.0, 114.8, 103.4, 96.9, 65.4, 61.4, 55.8, 55.5, 14.2; ESI-HRMS (m / z): C 20 H 22 O5 [M+H] + : calcd: 343.1467; found: 343.1529.

[0035] b. Intermediate 1 (12.00 g, 35.05 mmol) was weighed into a clean 250 mL round bottom flask, then 135 mL DMF was added, ice-salt bath to below 0°C, slowly dropwise added phosphorus oxychloride (8.06 g, 52.57 mmol), after dropwise addition, the reaction was stirred at room temperature, TLC monitored the reaction, after the reaction was completed, the reaction solution was slowly added into ice water, and the pH of the reaction solution was adjusted to weak alkaline with NaHC03, then filtered, the crude product was obtained, and column chromatography purification (DCM: PE = 3: 1) gave yellow-green solid, which was intermediate 2. 1 HNMR (400 MHz, CDC13) δ 7.44 (d, J = 8.6 Hz, 2H), 7.03 (d, J = 16.2 Hz, 1H), 6.94 - 6.88 (m, 3H), 6.67 - 6.62 (m, 2H), 6.38 (s, 1H), 4.63 (s, 2H), 4.28 (q, J = 7.1 Hz, 2H), 3.82 (s, 6H), 1.30 (t, J = 7.1 Hz, 3H). 13 C NMR (100 MHz, CDC13) δ 190.6, 168.8, 165.0, 164.5, 157.7, 142.9, 131.78, 131.0, 128.4, 126.1, 116.0, 114.8, 103.4, 96.9, 65.4, 61.4, 55.8, 55.5, 14.2; ESI-HRMS (m / z): C 21 H 22 O6 [M+H] +: calcd: 371.1416; found: 371.1490.

[0036] c. Intermediate 2 (6.60 g, 17.82 mmol) was added to a clean 100 mL round-bottom flask, followed by 20 mL of 10% ethanol, then 36 mL of 10% sodium hydroxide solution, and the reaction was stirred at 50 °C for 2 h. After the reaction was completed, 20 mL of ice water was added, and the pH was adjusted to slightly acidic with 3 mol / L HC1. Filtration was performed, and a dark yellow solid was obtained, which was intermediate 3; 1 H NMR (400 MHz, CDC13) δ 10.53 (s, 1H), 8.08 (d, J = 16.0 Hz, 1H), 7.53 (d, J = 8.7 Hz, 2H), 7.00 - 6.90 (m, 4H), 6.74 (s, 1H), 6.40 (s, 1H), 4.71 (s, 2H), 3.93 (s, 3H), 3.91 (s, 3H). 13 C NMR (100 MHz, DMSO-d6) δ 194.9, 175.3, 170.0, 169.7, 163.0, 146.9, 137.1, 135.3, 133.2, 129.8, 120.3, 120.0, 108.7, 102.6, 69.6, 61.4, 60.9; ESI-HRMS (m / z): C 20 H 22 O5[M+H] + : calcd: 343.1103; found: 343.1166.

[0037] d. Pynolidine (71.12 mg, 1.00 mmol) was added to a clean 25 mL round-bottom flask, followed by 1 mL of DMF, then intermediate 3 (0.20 g, 0.58 mmol), HATU (33.32 mg, 0.87 mmol), and DIPEA (11.33 mg, 0.87 mmol) were added in sequence. The reaction was stirred at room temperature, and TLC was used to monitor the reaction. After the reaction was completed, an appropriate amount of water was added dropwise to the reaction solution, and a solid was precipitated. Filtration was performed directly, the filter cake was recovered, and after drying, 380.1 mg of an orange-yellow solid was obtained, which was intermediate 4-1, with a yield of 91.14%. 1HNMR (400 MHz, CDC13) δ 10.53 (s, 1H), 8.05 (d, J = 16.2 Hz, 1H), 7.50 (d, J = 8.5 Hz, 2H), 6.99 - 6.92 (m, 3H), 6.74 (s, 1H), 6.40 (s, 1H), 4.65 (s, 2H), 3.92 (s, 3H), 3.90 (s, 3H), 3.54 (t, J = 6.6 Hz, 4H), 2.01 - 1.93 (m, 2H), 1.90 - 1.82 (m, 2H).13C NMR (100 MHz, CDC13) δ 166.4, 165.0, 164.5, 157.9, 143.1, 131.9, 130.8, 128.4, 126.1, 116.1, 114.8, 103.3, 97.0, 68.1, 55.9, 55.6, 46.3, 46.1, 26.3, 23.8. ESI-HRMS (m / z): C 23 H 25 NO5[M+H] + : calcd: 396.1766; found: 396.1794.

[0038] e. Intermediate 4-1 (197.6 mg, 0.5 mmol) was added to a clean 25 mL round bottom flask, then 1 mL of absolute ethanol was added, and then paeonol (124.7 mg, 0.75 mmol) and pyrrolidine (71.12 mg, 1 mmol) were added again. After stirring at 40 °C for 24 h, the reaction was concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (DCM:MeOH = 15:1) to obtain orange-yellow solid, which was compound C-1. 1 H NMR (600 MHz, CDC13) δ 13.63 (s, 1H), 8.23 (d, J = 15.4 Hz, 1H), 7.67 (d, J = 9.0 Hz, 1H), 7.60 (d, J = 15.4 Hz, 1H), 7.47 (d, J = 8.7 Hz, 2H), 7.34 (d, J = 16.0 Hz, 1H), 6.97 - 6.91 (m, 3H), 6.71 (d, J = 2.3 Hz, 1H), 6.45 (d, J = 2.4 Hz, 2H), 6.34 (dd, J = 9.0, 2.5 Hz, 1H), 4.64 (s, 2H), 3.92 (s, 3H), 3.90 (s, 3H), 3.83 (s, 3H), 3.53 (t, J = 6.8 Hz, 4H), 1.99 - 1.95 (m, 2H), 1.87 - 1.83 (m, 2H). 13CNMR (100 MHz, CDC13) δ 191.3, 165.2, 165.0, 164.5, 160.4, 159.5, 156.7, 140.4, 137.0, 130.3, 129.9, 129.2, 126.8, 124.6, 122.0, 114.6, 113.6, 113.0, 106.2, 102.5, 99.6, 96.4, 66.7, 54.5, 54.2, 54.2, 44.9, 44.7, 24.9, 22.5; ESI-HRMS (m / z): C 32 H 27 NO7[M+H] + : calcd: 544.2335; found: 544.2358.

[0039] Example 2: Preparation of (E)-3-(2,4-dimethoxy-6-((E)-4-(2-oxo-2-(piperidin-l- yl)ethoxy)styryl)phenyl)-l-(2-hydroxy-5-methoxyphenyl)prop-2-en-l-one (C-2)

[0040]

[0041] The preparation method is the same as Example 1, except that piperidine is used instead of pyrrolidine in step d to obtain intermediate 4-2, and then intermediate 4-2 is used instead of intermediate 4-1 in step e to obtain the target product (E)-3-(2,4-dimethoxy-6-((E)-4-(2-oxo-2-(piperidin-l- yl)ethoxy)styryl)phenyl)-l-(2-hydroxy-5-methoxyphenyl)prop-2-en-l-one. Orange yellow solid. 1 HNMR (600 MHz, CDC13) δ 13.63 (s, 1H), 8.23 (d, J = 15.4 Hz, 1H), 7.67 (d, J = 8.9 Hz, 1H), 7.60 (d, J = 15.5 Hz, 1H), 7.46 (d, J = 8.2 Hz, 2H), 7.33 (d, J = 15.7 Hz, 1H), 6.98 - 6.93 (m, 3H), 6.71 (d, J = 2.4 Hz, 1H), 6.44 (d, J = 2.4 Hz, 2H), 6.34 (dd, J = 8.9, 2.6 Hz, 1H), 4.69 (s, 2H), 3.91 (s, 3H), 3.90 (s, 3H), 3.83 (s, 3H), 3.41 (q, J = 7.1 Hz, 4H), 1.22 (t, J = 7.0 Hz, 3H), 1.14 (t, J = 7.1 Hz, 3H). 13C NMR (100 MHz, CDC13) δ 192.7, 166.9, 166.6, 165.9, 161.8, 160.9, 158.2, 141.8, 138.4, 131.7, 131.3, 130.6, 128.1, 126.0, 123.4, 116.0, 115.1, 114.4, 107.6, 103.9, 100.0, 97.8, 67.7, 55.9, 55.6, 55.6, 41.7, 40.4, 14.48, 12.9; ESI-HRMS (m / z): C 33 H 35 NO7[M+H] + : calcd: 573.2601; found: 573.2624.

[0042] Example 3: Preparation of (E)-3-(2,4-dimethoxy-6-((E)-4-(2-oxo-2-(piperazin-1- yl)ethoxy)styryl)phenyl)-1-(2-hydroxy-5-methoxyphenyl)prop-2-en-1-one (C-3)

[0043]

[0044] The preparation method is the same as Example 1, except that piperazine is used instead of pyrrolidine in step d to obtain intermediate 4-3, and then intermediate 4-3 is used instead of intermediate 4-1 in step e to obtain the target product (E)-3-(2,4-dimethoxy-6-((E)-4-(2-oxo-2-(piperazin-1-yl)ethoxy)styryl)phenyl)-1-(2-hydroxy-5-methoxyphenyl)prop-2-en-1-one, orange yellow solid. 1 HNMR (600 MHz, CDC13) δ 13.63 (s, 1H), 8.22 (d, J = 15.5 Hz, 1H), 7.89 (d, J = 8.8 Hz, 1H), 7.67 (d, J = 9.0 Hz, 1H), 7.47 (d, J = 8.4 Hz, 2H), 7.35 (d, J = 4.9 Hz, 1H), 6.96 - 6.93 (m, 3H), 6.71 (d, J = 2.3 Hz, 1H), 6.45 (d, J = 2.5 Hz, 2H), 6.34 (dd, J = 9.0, 2.6 Hz, 1H), 4.71 (s, 2H), 3.92 (s, 3H), 3.90 (s, 3H), 3.83 (s, 3H), 3.63 - 3.59 (m, 4H), 2.90 - 2.86 (m, 4H). 13C NMR (100 MHz, CDC13) δ 191.7, 165.6, 165.4, 164.9, 160.8, 159.9, 156.8, 140.8, 137.4, 130.7, 130.3, 129.8, 127.2, 125.1, 122.4, 115.0, 114.0, 113.4, 106.6, 102.9, 99.9, 96.8, 76.4, 76.1, 75.8, 66.7, 54.9, 54.6, 54.6, 45.1, 44.9, 44.4, 41.7; ESI-HRMS (m / z): C 32 H 34 N2O7[M+H] + : calcd: 559.2444; found: 559.2473.

[0045] Example 4: Preparation of (E)-3-(2,4-dimethoxy-6-((E)-4-(2-(4-methylpiperazin-1- yl)-2-oxoethoxy)styryl)phenyl)-1-(2-hydroxy-5-methoxyphenyl)prop-2-en-1-one (C-4)

[0046]

[0047] The preparation method is the same as Example 1, except that N-methylpiperazine is used instead of pyrrolidine in step d to obtain intermediate 4-4, and then intermediate 4-4 is used instead of intermediate 4-1 in step e to obtain the target product (E)-3-(2,4-dimethoxy-6-((E)-4-(2-(4-methylpiperazin-1-yl)-2-oxoethoxy)styryl)phenyl)-1-(2-hydroxy-5-methoxyphenyl)prop-2-en-1-one, orange yellow solid. 1 HNMR (600 MHz, CDC13) δ 13.63 (s, 1H), 8.22 (d, J = 15.5 Hz, 1H), 7.89 (d, J = 8.8 Hz, 1H), 7.67 (d, J = 9.0 Hz, 1H), 7.47 (d, J = 8.7 Hz, 2H), 7.28 (d, J = 8.5 Hz, 1H), 6.96 - 6.93 (m, 3H), 6.71 (d, J = 2.3 Hz, 1H), 6.45 (d, J = 2.4 Hz, 2H), 6.35 (dd, J = 8.9, 2.4 Hz, 1H), 4.71 (s, 2H), 3.92 (s, 3H), 3.90 (s, 3H), 3.83 (s, 3H), 3.78 (d, J = 8.4 Hz, 4H), 2.35 - 2.27 (m, 7H). 13C NMR (100 MHz, CDC13) δ 192.6, 166.5, 166.2, 165.8, 161.8, 160.9, 157.8, 141.8, 138.4, 131.6, 131.2, 130.7, 130.6, 128.2, 126.0, 123.4, 115.9, 115.0, 114.3, 114.2, 107.6, 103.8, 100.9, 97.7, 77.4, 77.1, 76.7, 67.6, 55.8, 55.6, 55.5, 55.1, 54.6, 46.0, 45.2, 42.0; ESI-HRMS (m / z): C 33 H 36 N2O7[M+H] + : calcd: 573.2601; found: 573.2624.

[0048] Example 5: Preparation of (E)-3-(2-((E)-4-(2-(4-ethylpiperazin-1-yl)-2- oxoe thoxy)styryl)-4,6-dimethoxyphenyl)-1-(2-hydroxy-5-methoxyphenyl)prop-2- en-1-one (C-5)

[0049]

[0050] The preparation method is the same as Example 1, except that N-ethylpiperazine is used to replace pyrrolidine in step d to obtain intermediate 4-5, and then intermediate 4-5 is used to replace intermediate 4-1 in step e to obtain the target product (E)-3-(2-((E)-4-(2-(4-ethylpiperazin-1-yl)-2-oxoethoxy)styryl)-4,6-dimethoxyphenyl)-1-(2-hydroxy-5-methoxyphenyl)prop-2-en-1-one, orange solid 185.9 mg, yield 63.44%. 1H NMR (600 MHz, CDC13) δ 13.63 (s, 1H), 8.22 (d, J = 15.5 Hz, 1H), 7.89 (d, J = 8.8 Hz, 1H), 7.67 (d, J = 9.0 Hz, 1H), 7.59 (d, J = 3.5 Hz, 1H), 7.46 (d, J = 5.6 Hz, 2H), 7.35 (d, J = 5.4 Hz, 1H), 6.95 - 6.93 (m, 2H), 6.71 (d, J = 2.3 Hz, 1H), 6.44 (d, J = 2.4 Hz, 2H), 6.34 (dd, J = 8.9, 2.5 Hz, 1H), 4.70 (s, 2H), 3.92 (s, 3H), 3.90 (s, 3H), 3.83 (s, 3H), 3.67 - 3.61 (m, 4H), 2.46 - 2.41 (m, 6H), 1.10 - 1.07 (m, 3H). 13 C NMR (100 MHz, CDC13) δ 191.6, 165.6, 165.2, 164.9, 160.8, 159.9, 156.9, 140.8, 137.4, 130.7, 130.2, 129.8, 127.2, 125.0, 122.4, 115.0, 114.0, 113.4, 106.6, 102.9, 100.0, 96.7, 76.4, 76.1, 75.8, 66.7, 54.9, 54.6, 51.9, 51.4, 51.29, 44.3, 41.1, 28.8; ESI-HRMS (m / z): C 34 H 38 N2O7[M+H] + : calcd: 587.2757; found: 587.2781.

[0051] Example 6: Preparation of (E)-3-(2-((E)-4-(2-(4-benzylpiperazin-l-yl)-2- oxoethoxy)styryl)-4,6-dimethoxyphenyl)-l-(2-hydroxy-5-methoxyphenyl)prop-2- en-l-one (C-6)

[0052]

[0053] The preparation method is the same as Example 1, except that N-benzylpiperazine is used instead of pyrrolidine in step d to obtain intermediate 4-6, and then intermediate 4-6 is used instead of intermediate 4-1 in step e to obtain the target product (E)-3-(2-((E)-4-(2-(4-benzylpiperazin-l-yl)-2-oxoethoxy)styryl)-4,6-dimethoxyphenyl)-l-(2-hydroxy-5-methoxyphenyl)prop-2-en-l-one, orange solid 203.4 mg, yield 62.76%.1 H NMR (600 MHz, CDC13) δ 13.64 (s, 1H), 8.23 (d, J = 15.5 Hz, 1H), 7.67 (d, J = 9.0 Hz, 1H), 7.61 (d, J = 15.5 Hz, 1H), 7.46 (d, J = 8.7 Hz, 2H), 7.38 - 7.25 (m, 7H), 6.95 - 6.91 (m, 2H), 6.72 (d, J = 2.3 Hz, 1H), 6.45 (t, J = 2.6 Hz, 2H), 6.34 (dd, J = 9.0, 2.5 Hz, 1H), 4.70 (s, 2H), 3.92 (s, 3H), 3.90 (s, 3H), 3.82 (s, 3H), 3.66 - 3.47 (m, 6H), 2.50 - 2.40 (m, 4H). 13 C NMR (100 MHz, CDC13) δ 192.7, 166.6, 166.2, 165.9, 161.8, 160.9, 157.9, 141.8, 138.4, 131.7, 131.3, 130.8, 129.2, 128.4, 128.2, 127.4, 126.1, 123.5, 116.0, 115.0, 114.4, 107.6, 103.9, 101.0, 97.8, 77.4, 77.1, 76.8, 67.8, 62.9, 55.9, 55.6, 55.6, 53.2, 52.7, 45.4, 42.2; ESI-HRMS (m / z): C 39 H 40 N2O7[M+H] + : calcd: 649.2914; found: 649.2933.

[0054] Example 7: Preparation of (E)-1-(2-hydroxy-5-methoxyphenyl)-3-(2-((E)-4-(2-(4-(2- hydroxyethyl)piperazin-1-yl)-2-oxoethoxy)styryl)-4,6-dimethoxyphenyl)prop-2-en-1-one (C-7)

[0055]

[0056] The preparation method is the same as that in Example 1, except that N-hydroxyethylpiperazine is used instead of pyrrolidine in step d to obtain intermediate 4-7, and then intermediate 4-7 is used instead of intermediate 4-1 in step e to obtain the target product (E)-1-(2-hydroxy-5-methoxyphenyl)-3-(2-((E)-4-(2-(4-(2-hydroxyethyl)piperazin-1-yl)-2-oxoethoxy)styryl)-4,6-dimethoxyphenyl)prop-2-en-1-one, an orange solid. 1H NMR (600 MHz, CDC13) δ 13.63 (s, 1H), 8.22 (d, J = 15.5 Hz, 1H), 7.67 (d, J = 9.0 Hz, 1H), 7.61 (d, J = 15.5 Hz, 1H), 7.47 (d, J = 8.3 Hz, 2H), 7.34 (d, J = 15.6 Hz, 2H), 6.95 - 6.92 (m, 2H), 6.71 (d, J = 2.3 Hz, 1H), 6.45 (d, J = 2.4 Hz, 2H), 6.35 (dd, J = 9.0, 2.5 Hz, 1H), 4.71 (s, 2H), 3.92 (s, 3H), 3.90 (s, 3H), 3.83 (s, 3H), 3.70 - 3.65 (m, 6H), 2.63-2.56 (m, 6H). 13 C NMR (100 MHz, CDC13) δ 192.74, 166.66, 166.37, 165.94, 161.9, 160.9, 157.8, 141.8, 138.4, 131.7, 131.3, 130.9, 128.2, 128.1, 126.17, 123.4, 116.0, 115.0, 114.9, 114.4, 107.7, 103.9, 101.0, 97.8, 77.4, 7.16, 76.8, 67.8, 59.4, 57.8, 55.9, 55.6, 55.6, 53.1, 52.6, 45.4, 42.1; ESI-HRMS (m / z): C 34 H 38 N2O8[M+H]+: calcd: 603.2706; found: 603.2738.

[0057] Example 8: Preparation of (E)-3-(2,4-dimethoxy-6-((E)-4-(2-morpholino-2-oxidoethoxy)styryl)phenyl)-1-(2-hydroxy-5-methoxyphenyl)prop-2-en-1-one (C-8)

[0058]

[0059] The preparation method is the same as Example 1, except that morpholine is used instead of pyrrolidine in step d to obtain intermediate 4-8, and then intermediate 4-8 is used instead of intermediate 4-1 in step e to obtain the target product (E)-3-(2,4-dimethoxy-6-((E)-4-(2-morpholino-2-oxidoethoxy)styryl)phenyl)-1-(2-hydroxy-5-methoxyphenyl)prop-2-en-1-one, orange solid. 1HNMR (600 MHz, CDC13) δ 13.63 (s, 1H), 8.22 (d, J = 15.5 Hz, 1H), 7.67 (d, J = 8.9 Hz, 1H), 7.60 (d, J = 15.4 Hz, 1H), 7.47 (d, J = 8.4 Hz, 2H), 7.34 (d, J = 15.9 Hz, 1H), 6.97 - 6.92 (m, 3H), 6.71 (d, J = 2.3 Hz, 1H), 6.45 (d, J = 2.5 Hz, 2H), 6.34 (dd, J = 8.9, 2.5 Hz, 1H), 4.72 (s, 2H), 3.92 (s, 3H), 3.90 (s, 3H), 3.83 (s, 3H), 3.67 (t, J = 4.7 Hz, 4H), 3.65 - 3.61 (m, 4H). 13 C NMR (100 MHz, CDC13) δ 192.7, 166.6, 166.5, 165.9, 161.8, 160.9, 157.7, 141.8, 138.4, 131.6, 131.3, 130.9, 128.2, 126.2, 123.4, 116.0, 115.0, 114.4, 107.6, 103.9, 101.0, 97.8, 77.4, 77.1, 76.8, 67.7, 66.9, 66.8, 55.9, 55.6, 55.6, 46.0, 42.5; ESI-HRMS (m / z): C 32 H 33 NO8 [M+H] + : calcd: 560.2284; found: 560.2240.

[0060] Example 9: Preparation of N,N-diethyl-2-(4-((E)-2-(E)-3-(2-hydroxy-5- methoxyphenyl)-3-oxopropyl-1-en-1-yl)-3,5-dimethoxystyryl)phenoxy)acetamide (C-9)

[0061]

[0062] The preparation method is the same as that in Example 1, except that diethylamine is used instead of pyrrolidine in step d to obtain intermediate 4-9, and then intermediate 4-9 is used instead of intermediate 4-1 in step e to obtain the target product N,N-diethyl-2-(4-((E)-2-(E)-3-(2-hydroxy-5-methoxyphenyl)-3-oxopropyl-1-en-1-yl)-3,5-dimethoxystyryl)phenoxy)acetamide, orange yellow solid. 1HNMR (400 MHz, CDC13) δ 13.69 (s, 1H), 8.26 (d, J = 15.5 Hz, 1H), 7.70 (d, J = 9.0 Hz, 1H), 7.63 (d, J = 15.5 Hz, 1H), 7.52 - 7.47 (m, 2H), 7.37 (d, J = 16.1 Hz, 1H), 7.00 - 6.96 (m, 3H), 6.74 (d, J = 2.3 Hz, 1H), 6.47 (dd, J = 2.5, 1.6 Hz, 2H), 6.38 (dd, J = 9.0, 2.5 Hz, 1H), 4.73 (s, 2H), 3.95 (s, 3H), 3.93 (s, 3H), 3.86 (s, 3H), 3.59 (t, J = 5.6 Hz, 2H), 3.52 (t, J = 5.4 Hz, 2H), 1.64 - 1.54 (m, 6H). 13 C NMR (100 MHz, CDC13) δ 192.7, 166.9, 166.6, 165.9, 161.8, 160.9, 158.2, 141.8, 138.4, 131.7, 131.3, 130.6, 128.1, 126.0, 123.4, 116.0, 115.1, 114.4, 107.6, 103.9, 101.0, 97.8, 77.4, 77.1, 76.8, 67.7, 55.9, 55.6, 55.6, 41.7, 40.4, 14.4, 12.9; ESI-HRMS (m / z): C 32 H 35 NO7 [M+H] + : calcd: 546.2492; found: 546.2518.

[0063] Example 10: Preparation of 2-(4-((E)-2-(E)-3-(2-hydroxy-5-methoxyphenyl)-3- oxopropyl- 1 -en- 1 -yl)-3,5 -dimethoxy styryl)phenoxy)-N-(2-hydroxyethyl)acetamide (C-10)

[0064]

[0065] The preparation method is the same as Example 1, except that ethanolamine is used instead of pyrrolidine in step d to obtain intermediate 4-10, and then intermediate 4-10 is used instead of intermediate 4-1 in step e to obtain the target product 2-(4-((E)-2-(E)-3-(2-hydroxy-5-methoxyphenyl)-3-oxopropyl- 1 -en- 1 -yl)-3,5 -dimethoxy styryl)phenoxy)-N-(2-hydroxyethyl)acetamide, orange yellow solid 165.6 mg, yield 62.12%. 1H NMR (400 MHz, CDC13) δ 8.24 (d, J = 15.5 Hz, 1H), 7.68 (d, J = 9.0 Hz, 1H), 7.62 (d, J = 15.5 Hz, 1H), 7.51 (d, J = 8.6 Hz, 2H), 7.37 (d, J = 16.0 Hz, 1H), 6.97 - 6.91 (m, 3H), 6.72 (d, J = 1.9 Hz, 1H), 6.46 (d, J = 2.0 Hz, 2H), 6.35 (dd, J = 8.9, 2.4 Hz, 1H), 4.55 (s, 2H), 3.93 (s, 3H), 3.91 (s, 3H), 3.84 (s, 3H), 3.82 - 3.78 (t, J = 4.8 Hz, 2H), 3.59 - 3.54 (t, J = 5.4 Hz, 2H). 13 C NMR (100 MHz, CDC13) δ 191.6, 165.5, 164.9, 164.8, 160.8, 159.8, 157.0, 140.8, 137.4, 130.7, 130.2, 129.5, 127.1, 124.9, 122.4, 114.9, 113.9, 113.3, 106.5, 102.8, 99.9, 96.7, 76.3, 76.0, 75.7, 66.7, 54.8, 54.5, 54.5, 45.5, 42.2, 25.5, 24.5, 23.4; ESI-HRMS (m / z): C 30 H 31 NO8[M+H] + : calcd: 534.2128; found: 534.2141.

[0066] Example 11: Evaluation of Anti-tumor Activity

[0067] 1. CCK-8 method for determining in vitro anti-tumor activity

[0068] The CCK-8 (Cell Counting Kit-8) method was used to determine the inhibition rate of the pterostilbene derivatives on human lung cancer cells A549, human liver cancer cells HepG2, colon cancer cells HCT116, and human osteosarcoma cells U-2OS.

[0069] Four kinds of human tumor cells were stored in RPMI-1640 medium containing 10% fetal bovine serum (BI), 100 U / mL of penicillin and 0.1 mg / mL of streptomycin at 37°C in a 5% CO2 cell incubator; when the cells were in the logarithmic phase, the cells were collected and counted, diluted to an appropriate density, inoculated into a 96-well plate, the inoculation number of adherent cells was 5000 / well, the inoculation number of suspended cells was 8000 / well, and the edge wells were covered with PBS, and placed in a culture incubator (condition: 37°C, 5% CO2) overnight; the prepared 100 μM concentration of the test compound and the positive drug stock solution were diluted according to the gradient dilution method to prepare different concentrations of the test solution with the cell culture solution. 100 μL of the test solution was added to each well, 5 concentrations of each compound were set, and 3 replicate wells were set for each concentration. The blank group was without cells, the negative control group was without drugs, and the same volume of culture medium was used as a control. After adding the drugs, the medium was returned to the incubator and cultured at 37°C in a 5% CO2 cell incubator for 48 h. Then 10 μL of CCK-8 solution was added to each well, and incubated for about 2 h under the same conditions. Finally, the absorbance (OD value) at 450 nm was determined by an enzyme-labeled instrument. Each group of experiments was repeated three times, and each experiment was independent.

[0070] The inhibition rate of cell growth was calculated according to the following formula:

[0071] The growth inhibition rate = (1-survival rate) x 100% = [1-(OD 实验 -OD 空 ) / (OD 对照 -OD 空 )] x 100% (OD experimental represents the absorbance of the test drug group, OD 对照 represents the absorbance of the control group, and OD 空白 represents the absorbance of the blank group).

[0072] The half maximal inhibitory concentration (IC 50 ) is defined as the drug concentration when 50% of the tumor cells survive. According to the measured absorbance (OD value), the inhibition rate was calculated, and then the IC 50 value was fitted using Origin.

[0073] The measured IC 50 values are shown in Table 1.

[0074] Table 1 IC 50 values of the listed pterostilbene compounds of the present application on four kinds of tumor cells

[0075]

[0076] From Table 1, it can be seen that the inhibitory activities of most of the compounds on A549, HepG2, HCT116 and U-2OS tumor cells are stronger than that of pterostilbene, and the activities of some of the compounds are stronger than that of the HDAC inhibitor chidamide.

[0077] 2. HDAC inhibition experiment method

[0078] The adherent cells A549 in the logarithmic growth phase were taken, and the cell suspension density was adjusted to 1 x 10 6 / mL. The cells were inoculated in a six-well plate, and different concentration administration groups and a negative control group were set. After 24 h of cell inoculation and complete cell adhesion, the culture medium was aspirated, and the cells were cultured for 24 h after administration. After the culture medium was aspirated and washed with PBS, the adherent cells were digested into a single cell suspension with trypsin, and then the culture medium was added to terminate the digestion. Subsequently, the cell solution was added to an EP tube, centrifuged to discard the supernatant, washed once with pre-cooled PBS, and finally 100 μL of lysis buffer was added to ultrasonically break the cells for 30 s in an ultrasonic breaker. The supernatant was obtained by centrifugation. The intracellular protein sample was stored at -20 °C.

[0079] The human histone deacetylase enzyme-linked immunosorbent assay kit was used, and blank wells, negative control wells, standard wells and administration wells were set. After incubation, the OD value at 450 nm was measured using an enzyme marker, a standard curve was obtained according to the standard, and the HDAC concentration of the different concentration administration groups was calculated. Each group of experiments was repeated three times, and each experiment was independent.

[0080]

[0081] After the inhibition rate of the compounds at different concentrations was calculated using the formula, the Origin software was used to fit the inhibition curve, and the IC 50 value was obtained.

[0082] According to the in vitro anti-tumor experiment, three best active compounds, C-2, C-3 and C-4, were taken. The three compounds were subjected to in vitro HDAC enzyme inhibition experiment, and the experimental results are shown in Table 2.

[0083] Table 2 HDAC inhibition activity of compounds C-2 to 4

[0084]

[0085] a The mean value ± standard deviation of three experiments is represented

[0086] The in vitro HDAC inhibition activity experiment results show that the three compounds have certain inhibition activity on five kinds of HDAC enzymes, and the inhibition activity of the three target compounds on HDAC1, 2 and 3 is better than that on HDAC6 and 8.

[0087] The foregoing is considered as illustrative only of the principles of the application and the forms thereof shown and described by way of example. Further, those skilled in the art will realize that the mechanism of the present application is capable of other modes of practice and still be within the spirit and scope of the application. For example, the application can be used in other types of systems and / or methods. The disclosure should be understood, therefore, as not being so limited, but rather directed primarily to the principal of the application and the major features thereof. Further, changes and modifications can be obvious to those skilled in the art, and are intended to be included within the spirit and scope of the application as defined by the following claims.

Claims

1. Pterostilbene paeonol chalcone amide derivatives represented by general formula (I) or pharmaceutically acceptable salts thereof: Formula (I); in, R is selected from piperidinyl, morpholinyl, piperazinyl, N -methylpiperazinyl, N -ethylpiperazinyl, N -Benzylpiperazinyl, N -Any one of a hydroxyethylpiperazinyl group, a tetrahydropyrrolyl group, a C1-C6 alkylamino group or a 2-hydroxyethylamino group.

2. A method for preparing the pterostilbene paeonol chalcone amide derivatives according to claim 1, characterized in that: The following steps are involved: S1. Under alkaline conditions, pterostilbene, the compound represented by formula (1) and a solvent are contacted to perform a nucleophilic substitution reaction to obtain a compound represented by formula (2). The reaction formula is as follows: ; wherein X is a halogen; S2. The compound represented by formula (2), a formylating agent, and an acidic chloride are contacted to perform a Vilsmeier-Haack reaction to obtain a compound represented by formula (3). The reaction formula is as follows: S3. Under alkaline conditions, the compound represented by formula (3) is contacted with a solvent for hydrolysis reaction to obtain a compound represented by formula (4). The reaction formula is as follows: ； S4. The compound represented by formula (4), compound (5), a condensing agent and a solvent are contacted to perform a nucleophilic substitution reaction to obtain a compound represented by formula (6). The reaction formula is as follows: ； The compound (5) is selected from piperidine, morpholine, piperazine, N -Methylpiperazine, N -ethylpiperazine, N -Benzylpiperazine, N -Hydroxyethylpiperazine, pyrrolidine, C1-C6 alkylamine compounds, 2-hydroxyethylamine; R is defined as in claim 1; S5. Under alkaline conditions, the compound represented by formula (6), paeonol and a solvent are contacted to carry out an aldol condensation reaction to obtain a compound represented by formula (I), as shown in the following reaction formula:

3. The preparation method according to claim 2, characterized in that In step S1, the molar ratio of pterostilbene to the compound represented by formula (1) is 1:1-3.

4. The preparation method according to claim 3, characterized in that In step S1, the base used in the alkaline condition is potassium carbonate, and the solvent is acetonitrile.

5. The preparation method according to claim 3, characterized in that In step S1, the reaction temperature is 60-90° C., and the reaction time is 12-24 h.

6. The preparation method according to claim 2, characterized in that In step S2, the formylating agent is N,N -dimethylformamide.

7. The preparation method according to claim 2, characterized in that In step S2, the reaction temperature is room temperature and the reaction time is 2 to 4 hours.

8. The preparation method according to claim 2, characterized in that In step S3, the base used in the alkaline condition is sodium hydroxide solution, and the solvent is ethanol.

9. The preparation method according to claim 8, characterized in that In step S3, the reaction temperature is 40-60° C., and the reaction time is 1-4 h.

10. The preparation method according to claim 2, characterized in that In step S4, the molar ratio of the compound represented by formula (4) to compound (5) is 1:1-3.

11. The preparation method according to claim 10, characterized in that: In step S4, the condensing agent is HATU, DIPEA, and the solvent is N,N -dimethylformamide.

12. The preparation method according to claim 10, characterized in that In step S4, the reaction temperature is room temperature, and the reaction time is 1 to 3 hours.

13. The preparation method according to claim 2, characterized in that In step S5, the molar ratio of the compound represented by formula (6) to paeonol is 1:1-3.

14. The preparation method according to claim 13, characterized in that In step S5, the base used in the alkaline condition is pyrrolidine, and the solvent is ethanol.

15. The preparation method according to claim 13, characterized in that In step S5, the reaction temperature is 30-60° C., and the reaction time is 12-24 h.

16. A pharmaceutical composition, characterized in that: The invention is composed of a therapeutically effective amount of the pterostilbene paeonol chalcone amide derivatives or a pharmaceutically acceptable salt thereof as claimed in claim 1 and a pharmaceutically acceptable carrier.

17. The pharmaceutical composition according to claim 16, characterized in that The dosage form of the pharmaceutical composition is any one of ordinary tablets or capsules, sustained-release tablets or capsules, controlled-release tablets or capsules, oral solution, and injection.

18. Use of the pterostilbene paeonol chalcone amide derivatives or pharmaceutically acceptable salts thereof according to claim 1 in the preparation of anti-tumor drugs, characterized in that: When R is tetrahydropyrrolyl, the tumor is colon cancer or lung cancer; When R is piperidinyl or piperazinyl, the tumor is liver cancer, colon cancer, human osteosarcoma or lung cancer; When R is N -methylpiperazine, the tumor is liver cancer, colon cancer or human osteosarcoma; When R is N -benzylpiperazinyl, the tumor is colon cancer, human osteosarcoma or lung cancer; When R is N -ethylpiperazinyl, N -hydroxyethylpiperazinyl, C1-C6 alkylamino, the tumor is lung cancer; When R is a morpholinyl group, the tumor is colon cancer or lung cancer; When R is 2-hydroxyethylamino, the tumor is liver cancer.

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