Resveratrol derivative as well as preparation method and application thereof

Abstract

The invention discloses a resveratrol derivative and a preparation method and application thereof, and relates to the field of organic synthesis, the structural general formula of the resveratrol derivative is as shown in the following formula 1: # imgabs0 # R1 and R2 both comprise one of # imgabs1 # imgabs2 #, and the two are the same or different; wherein the # imgabs3 # is a linking site. According to the application, resveratrol is chemically modified to obtain the resveratrol derivative containing R1 and R2 groups, so that the inhibition effect on key proteins such as mTOR (mammalian Target of Radical), JAK (Janus Kinase), beta-amyloid, Adenyl cyclase and IKK beta is improved, and the anti-tumor activity of the resveratrol derivative is improved.

Description

Technical Field

[0001] This application relates to the field of organic synthesis, and particularly to a resveratrol derivative, a preparation method thereof, and an application thereof. Background Art

[0002] As a naturally occurring polyphenolic compound, resveratrol is widely present in grape skins, red wine, peanuts, and some plants, and has received extensive attention due to its remarkable antioxidant, anti-inflammatory, and potential anti-cancer properties. However, the stability of resveratrol in vivo is poor and its bioavailability is low, which limits its application potential in the medical field.

[0003] Currently, the research on resveratrol focuses on synthesizing resveratrol derivatives through esterification, etherification, oxidation, and binding with amino acids or sugar molecules. However, there is still a problem of weak inhibitory activity against the in vitro proliferation of tumor cells such as gastric cancer SGC-7901, lung cancer H460, liver cancer HepG2, and colorectal cancer CT26. Summary of the Invention

[0004] In view of the deficiencies of the above-mentioned related technologies, this application provides a resveratrol derivative, a preparation method thereof, and an application thereof. By chemically modifying resveratrol, a resveratrol derivative containing R 1 and R 2 groups is obtained, which improves the inhibitory effects on key proteins such as mTOR, JAK, β-amyloid, Adenylyl cyclase, and IKKβ, and enhances the anti-tumor activity of the resveratrol derivative.

[0005] In a first aspect, a resveratrol derivative provided by this application adopts the following technical solution:

[0006] A resveratrol derivative has the following general structural formula as shown in Formula 1:

[0007]

[0008] R 1 and R 2 both include one of them, and they are the same or different;

[0009] where is the linking site.

[0010] Preferably, R 1 and R 2 are the same.

[0011] Preferably, its structural formula includes one of the following structural formulas:

[0012]

[0013] In a second aspect, a preparation method of a resveratrol derivative provided by the present application adopts the following technical solution:

[0014] It includes the following steps: mixing resveratrol with one or both of a halogen and a hydrogen halide for an addition reaction, oxidizing the excess halogen and / or hydrogen halide and then adjusting the pH to 6-7, spin-drying and purifying to obtain a halogenated resveratrol; mixing the halogenated resveratrol, a reactant, a catalyst, and a solvent, stirring and reacting in an inert gas environment at 80-100 °C for 11-13 h, then cooling to room temperature, diluting the reaction system and extracting, retaining the organic phase, drying and concentrating the organic phase to a paste and then purifying to obtain the resveratrol derivative.

[0015] Preferably, the reactant includes one or two of the following.

[0016] Preferably, the catalyst includes one or more of a palladium catalyst and a basic substance.

[0017] Preferably, the palladium catalyst includes one or more of palladium on carbon, tetrakis(triphenylphosphine)palladium, palladium acetate, dichlorobis(η5-cyclopentadienyl)iron(II) diphenylphosphine, and dichlorobis(triphenylphosphine)palladium.

[0018] Preferably, the basic substance includes one or more of sodium carbonate, sodium bicarbonate, potassium carbonate, and potassium bicarbonate.

[0019] Preferably, the catalyst includes a palladium catalyst and a basic substance with a molar equivalent ratio of 1:145-155.

[0020] Preferably, the catalyst includes a palladium catalyst and a basic substance with a molar equivalent ratio of 1:150.

[0021] Preferably, the catalyst includes palladium on carbon and sodium carbonate with a molar equivalent ratio of 1:150.

[0022] Preferably, the step of mixing resveratrol with one or both of a halogen and a hydrogen halide for an addition reaction includes the following steps: dispersing resveratrol in a solvent and controlling the temperature at 75-100 °C, and dropping the hydrogen halide for 25-40 min.

[0023] Preferably, the step of mixing resveratrol with one or both of a halogen and a hydrogen halide for an addition reaction includes the following steps: dispersing resveratrol in a solvent and controlling the temperature at 80 °C, and dropping the hydrogen halide for 30 min.

[0024] Preferably, the molar ratio of resveratrol to hydrogen halide is 1:2-2.2.

[0025] In a third aspect, the present application provides an application of a resveratrol derivative in the preparation of a drug for inhibiting the proliferation of rectal cancer cells, gastric cancer cells, lung cancer cells and liver cancer cells.

[0026] Preferably, the resveratrol derivative is used as the main drug in tablets, capsules, granules, pills, syrups, oral solutions, oral suspensions and oral emulsions.

[0027] Preferably, the injection preparation includes injection solutions and powder for injection.

[0028] Preferably, the resveratrol derivative is used as an active ingredient of one or more of an mTOR inhibitor, a JAK inhibitor, a β-amyloid inhibitor, an Adenylyl cyclase inhibitor and an IKKβ inhibitor.

[0029] Preferably, the dosage of the resveratrol derivative is 4 - 50 μmol / L.

[0030] In summary, the present application includes at least one of the following beneficial technical effects:

[0031] 1. In the present application, a resveratrol derivative containing R 1 and R 2 groups is obtained by chemically modifying resveratrol, which improves the inhibitory effects on key proteins such as mTOR, JAK, β-amyloid, Adenylyl cyclase and IKKβ, and enhances the anti-tumor activity of the resveratrol derivative.

[0032] 2. The average increase in the in vitro proliferation inhibitory activity of the resveratrol derivative of the present application compared to resveratrol against gastric cancer SGC-7901, lung cancer H460, liver cancer HepG2 and colorectal cancer CT26 exceeds 20%.

[0033] 3. The HOMO-LUMO energy level gap of the resveratrol derivative of the present application is relatively short, between 2.44 eV and 2.58 eV, which is smaller than the band gap (3.18 eV) of the original resveratrol. This reduces the difficulty of the resveratrol derivative participating in electron transfer in chemical reactions or biological processes, is beneficial to its metabolism and function in the body, and can more easily transfer electrons to neutralize free radicals as a free radical scavenger, thereby enhancing its antioxidant ability. Detailed implementation manners

[0034] The following further elaborates on the present application in conjunction with embodiments. The following embodiments are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention. For those conditions not specified in the following embodiments, they are carried out according to conventional conditions or the conditions recommended by the manufacturer. The methods used, unless otherwise specified, are all conventional methods well-known in the art. The consumables and reagents used, unless otherwise specified, are all commercially available. Unless otherwise stated, the professional and scientific terms used herein have the same meaning as those familiar to skilled personnel in the art. In addition, any method or material similar or equivalent to the described content can also be applied to the present invention.

[0035] The raw materials used in the examples and comparative examples are all commercially available.

[0036] The synthesis process in the preparation of the embodiments of the present application is as follows:

[0037]

[0038] Example 1

[0039] Example 1 of the present application provides a resveratrol derivative, which is prepared by the following steps:

[0040] S1. Add concentrated HBr (mass concentration is 48%, total amount of HBr is 0.02 mol) dropwise to a mixture of resveratrol (0.01 mol) and ethanol (20 mL) at 80 °C for 30 min, and then add H 2 O 2 (total amount of H 2 O 2 is 0.02 mol). During this period, the color of the solution turns brown (the characteristic color of bromine). Continue stirring until the color disappears. Wait until it cools to room temperature, and further neutralize the solution by adding an aqueous solution of NaHCO 3 until the pH value is 6. Rotavapor to dryness, purify by passing through a silica gel column to obtain halogenated resveratrol (5-(1,2-dibromo-2-(4-hydroxyphenyl)ethyl)benzene-1,3-diol);

[0041] S2. Add halogenated resveratrol (0.01 mol), reactant (0.01 mol), palladium on carbon (0.01 eq), and sodium carbonate (1.5 eq) to 20 ml of dry dichloromethane solution. Under nitrogen protection, stir the reaction system at 85 °C for 4 hours, and then cool to room temperature. Dilute the reaction system with water, extract with dichloromethane, retain the organic phase, dry the organic phase with anhydrous magnesium sulfate, concentrate the organic phase to a paste, add 50 ml of ethanol, stir under reflux for 2 h, filter while hot, retain the organic phase, rotavapor the organic phase to dryness, purify by passing through a silica gel column to obtain a resveratrol derivative, named resveratrol derivative a. The structural formula of the target product is:

[0042]

[0043] In this embodiment, the reactants are:

[0044] The 1H NMR spectrum of haloresveratrol was detected, and the results are as follows: 1HNMR(400 MHz, chloroform-d): δ 8.38 (s, 2H), 7.32 - 7.26 (m, 2H), 7.14 (m, 1H), 6.82 - 6.76 (m, 2H), 6.56 - 6.52 (m, 2H), 6.17 (t, 1H), 5.73 - 5.67 (m, 1H), 5.59 - 5.54 (m, 1H).

[0045] Example 2

[0046] Embodiment 2 of this application provides a resveratrol derivative. The difference between Embodiment 2 and Embodiment 1 is that the reactants used in step S2 of Embodiment 2 are: A resveratrol derivative was obtained, named resveratrol derivative b, and the structural formula of the target product is:

[0047] Example 3

[0048] Embodiment 3 of this application provides a resveratrol derivative. The difference between Embodiment 3 and Embodiment 1 is that the reactants used in step S2 of Embodiment 3 are: A resveratrol derivative was obtained, named resveratrol derivative c, and the structural formula of the target product is:

[0049] Example 4

[0050] Embodiment 4 of this application provides a resveratrol derivative. The difference between Embodiment 4 and Embodiment 1 is that the reactants used in step S2 of Embodiment 4 are: A resveratrol derivative was obtained, named resveratrol derivative d, and the structural formula of the target product is:

[0051] Example 5

[0052] Embodiment 5 of this application provides a resveratrol derivative. The difference between Embodiment 5 and Embodiment 1 is that the reactants used in step S2 of Embodiment 5 are: A resveratrol derivative was obtained, named resveratrol derivative e, and the structural formula of the target product is:

[0053] Example 6

[0054] Example 6 of this application provides a resveratrol derivative. The difference between Example 6 and Example 1 is that the reactants used in step S2 of Example 6 are: A resveratrol derivative was obtained, named resveratrol derivative f, and the structural formula of the target product is:

[0055] Example 7

[0056] Example 7 of this application provides a resveratrol derivative. The difference between Example 7 and Example 1 is that the reactants used in step S2 of Example 7 are: A resveratrol derivative was obtained, named resveratrol derivative g, and the structural formula of the target product is:

[0057] Example 8

[0058] Example 8 of this application provides a resveratrol derivative. The difference between Example 8 and Example 1 is that the reactants used in step S2 of Example 8 are: A resveratrol derivative was obtained, named resveratrol derivative h, and the structural formula of the target product is:

[0059] Example 9

[0060] Example 9 of this application provides a resveratrol derivative. The difference between Example 9 and Example 1 is that the reactants used in step S2 of Example 9 are: A resveratrol derivative was obtained, named resveratrol derivative i, and the structural formula of the target product is:

[0061] Test detection

[0062] (1) The 1H NMR, MS and yields of the resveratrol derivatives a - i in Examples 1 - 9 are shown in Table 1 below.

[0063] Table 1:

[0064]

[0065]

[0066] (2) The resveratrol derivatives of Examples 1-9 and the positive control drug (resveratrol) were each completely dissolved in DMSO and diluted to a concentration of 5 μmol / L with serum-free medium to obtain the test solutions of the resveratrol derivatives of Examples 1-9 and the test solution of the positive control drug. The MTT method was used to test the in vitro proliferation inhibitory activities of the test solutions of the resveratrol derivatives of Examples 1-9 and the test solution of the positive control drug against four tumor cell lines (SGC-7901, H460, HepG2, and CT26). The steps were as follows: (1) Four of the above-mentioned tumor cell lines in the logarithmic growth phase were each taken and inoculated into a 96-well plate, cultured for 24 h, and then 100 μL of one of the test solutions of the resveratrol derivatives of Examples 1-9 and the test solution of the positive control drug was added to the wells. Three parallel wells were set in each group and cultured in an incubator for another 36 h; (2) Then 20 μL of MTT solution with a mass concentration of 5 mg / mL was added and cultured for another 24 h, centrifuged, the supernatant was discarded, 150 μL of DMSO was added, and it was shaken at low speed in the dark until completely dissolved; (3) The optical density (OD) value was measured with an enzyme-linked immunosorbent assay (ELISA) reader at a wavelength of 570 nm, and the inhibition rate was calculated. The results are shown in Table 2.

[0067] Table 2:

[0068]

[0069] (3) The combination of resveratrol and the resveratrol derivatives of Examples 1-9 with mTOR, JAK, β-amyloid, Adenylyl cyclase, and IKKβ proteins was verified by molecular docking. The binding force between the two was negatively correlated with the magnitude of the binding energy. The protein structures were from the Protein Data Bank (PDB); the hydrogenation treatment of the protein molecules was completed by AutoDockTools to generate PDBQT files and determine the active pockets; the docking binding energy was calculated with Vina software; the visualization of the optimal binding model was constructed with PyMOL software. The results are shown in Table 3.

[0070] Table 3:

[0071]

[0072]

[0073] (4) The density functional DFT / B3LYP method in the Gaussian 16 program was used to optimize the geometric configurations of the compounds with 6-31G(d) as the basis set; the DFT-D3 with BJ-damping was used to correct the weak interactions to improve the calculation accuracy; finally, the HOMO and LUMO energy level diagrams of resveratrol and the resveratrol derivatives of Examples 1-9 were obtained, and their band gap values were calculated through calculation. The results are shown in Table 4 below.

[0074] Table 4:

[0075]

[0076] Result Analysis

[0077] The present application will be described in detail below in combination with the experimental results provided in Tables 1-4.

[0078] As can be seen from Table 1, the resveratrol derivatives prepared in Examples 1-9 of the present application are the target products, and the yields reach 52.3%-64.1%.

[0079] As can be seen from Table 2, the in vitro proliferation inhibitory activities of the resveratrol derivatives in Examples 1-9 of the present application against four tumor cell lines (SGC-7901, H460, HepG2, and CT26) are significantly improved compared with the positive control resveratrol. The in vitro proliferation inhibitory activity of the resveratrol derivatives in Examples 1-9 of the present application against SGC-7901 is increased by 24%-60% compared with the in vitro proliferation inhibitory activity of the positive control resveratrol against SGC-7901. The in vitro proliferation inhibitory activity of the resveratrol derivatives in Examples 1-9 of the present application against H460 is increased by 24.4%-64.4% compared with the in vitro proliferation inhibitory activity of the positive control resveratrol against H460. The in vitro proliferation inhibitory activity of the resveratrol derivatives in Examples 1-9 of the present application against HepG2 is increased by 12.7%-45.5% compared with the in vitro proliferation inhibitory activity of the positive control resveratrol against HepG2. The in vitro proliferation inhibitory activity of the resveratrol derivatives in Examples 1-9 of the present application against CT26 is increased by 17.3%-44.2% compared with the in vitro proliferation inhibitory activity of the positive control resveratrol against SGC-7901. Among them, the resveratrol derivatives in Examples 2, 5, and 9 are particularly prominent in the inhibition of gastric cancer cell SGC-7901, and the highest inhibition rate reaches 42.0%.

[0080] As can be seen from Table 3, the binding affinities of the resveratrol derivatives in Examples 1-9 of the present application with mTOR, JAK, β-amyloid, Adenylyl cyclase, and IKKβ proteins are significantly improved compared with resveratrol, indicating that the resveratrol derivatives in Examples 1-9 of the present application significantly improve the interaction with the above protein targets compared with resveratrol, and the pharmacological activities are significantly improved.

[0081] As can be seen from Table 4, resveratrol itself has a relatively high HOMO value (-7.42 eV) and a relatively low LUMO value (-4.24 eV), with a band gap (3.18 eV), and it is not prone to charge transfer reactions. The LUMO and HOMO values of the resveratrol derivatives in Examples 1-9 are close to each other, and are closer to 0 compared to resveratrol. The molecular orbital energies are generally low. It is analyzed that due to the introduction of large conjugated functional groups in the resveratrol derivatives of Examples 1-9, the spatial configuration is changed, resulting in a change in the electron cloud distribution, thereby reducing the energy level of the molecular orbitals. Moreover, the band gap range of the resveratrol derivatives in Examples 1-9 from 2.44 eV to 2.58 eV is much smaller than that of resveratrol, indicating that the resveratrol derivatives in Examples 1-9 are more likely to participate in electron transfer in chemical reactions or biological processes.

[0082] This specific embodiment is only an explanation of the present application, and it does not limit the present application. Those skilled in the art can make modifications to this embodiment without creative contributions according to needs after reading this specification, but as long as it is within the scope of the claims of the present application, it is protected by the patent law.

Claims

1. A resveratrol derivative, characterized in that: Its general structural formula is shown in Formula 1 below: Both R1 and R2 include One of the following, and the two are the same or different; in The linking site.

2. A resveratrol derivative according to claim 1, characterized in that: Its structural formula includes one of the following structural formulas:

3. A method for preparing the resveratrol derivative according to claim 1 or 2, characterized in that: The following steps are involved: One or both of halogen and hydrogen halide are mixed with resveratrol for addition reaction, excess halogen and / or hydrogen halide are oxidized and the pH is adjusted to 6-7, and the mixture is spin-dried and purified to obtain halogenated resveratrol; halogenated resveratrol, reactants, catalyst and solvent are mixed, stirred and reacted at 80-100° C. for 11-13 hours in an inert gas environment, and then cooled to room temperature, the reaction system is diluted and extracted, the organic phase is retained, and the organic phase is dried and concentrated to a muddy state and then purified to obtain the resveratrol derivative.

4. The method for preparing a resveratrol derivative according to claim 3, characterized in that: The reactants include One or both of the following.

5. The method for preparing a resveratrol derivative according to claim 3, characterized in that: The catalyst includes one or more of a palladium catalyst and an alkaline substance.

6. The method for preparing a resveratrol derivative according to claim 3, characterized in that: The catalyst comprises a palladium catalyst and an alkaline substance in a molar equivalent ratio of 1:145-155.

7. The method for preparing a resveratrol derivative according to claim 3, characterized in that: The addition reaction of using one or both of halogen and hydrogen halide mixed with resveratrol comprises the following steps: dispersing resveratrol in a solvent and controlling the temperature to be 75-100° C., and dripping hydrogen halide for 25-40 minutes.

8. The method for preparing a resveratrol derivative according to claim 7, characterized in that: The molar ratio of resveratrol to hydrogen halide is 1:2-2.

2.

9. Use of the resveratrol derivative according to claim 1 or 2 in the preparation of a drug for inhibiting the proliferation of colorectal cancer cells, gastric cancer cells, lung cancer cells and liver cancer cells.

10. The use of a resveratrol derivative according to claim 9, characterized in that: The resveratrol derivative is used as one or more effective ingredients of an mTOR inhibitor, a JAK inhibitor, a β-amyloid inhibitor, an Adenylyl cyclase inhibitor and an IKKβ inhibitor.

Images