Magnolia officinalis derivatives and their preparation methods and applications

Abstract

The present invention relates to a magnolol derivative and a preparation method and application thereof. The magnolol derivative comprises: C 37 H 46 O6 (GYS17). This invention achieves the first chemical splicing of magnolol and idebenone through chemical methods, resulting in a novel class of magnolol derivatives. Biological evaluation revealed that the magnolol derivative GYS17 significantly inhibits H2O2 oxidative damage in myocardial H9C2 cells, with significantly greater efficacy than the positive controls salvianolate and idebenone. The synthesis of this magnolol derivative has important theoretical and practical implications for the development of novel anti-myocardial injury drugs. This technology also provides new insights and approaches for the structural modification of magnolol derivatives.

Description

Technical Field

[0001] The present invention relates to the technical field of medicines, and in particular to a magnolol derivative and a preparation method and application thereof. Background Art

[0002] Myocardial injury is a major complication after surgery for patients with cancers such as breast cancer and pancreatic cancer. Studies have shown that the mortality rate of breast cancer patients due to chemotherapy complications, particularly cardiac factors, has increased significantly in recent years. Myocardial injury caused by cancer treatment has become a major issue that needs to be addressed in cancer chemotherapy research.

[0003] Using traditional Chinese medicine monomers or their backbones as combinatorial chemistry templates to design and synthesize novel active molecules has proven to be a highly effective approach to drug discovery. Magnolol, a class of biphenyldiphenol compounds isolated from the traditional Chinese medicine Magnolia officinalis, exhibits diverse activities, including antimyocardial damage, antitumor, and antioxidant activities. In recent years, research on the synthesis and structural modification of magnolol and its derivatives has become a hot topic.

[0004] However, the reported chemical modification studies on compounds containing hydroxyl functional groups, such as magnolol, have mainly focused on the conventional transformation method of esterification between the hydroxyl groups in the compound and the carboxyl groups in carboxylic acids. The structural modification of magnolol faces the following difficulties: (1) The phenolic hydroxyl group of magnolol is affected by the steric hindrance of the biphenol structure, making magnolol less reactive than monocyclic phenol compounds and less likely to undergo nucleophilic substitution reactions at the phenolic hydroxyl site; (2) magnolol contains two symmetrical phenolic hydroxyl groups, and further optimization of the conditions is required to ensure that other raw materials react with only one of the hydroxyl groups to obtain the target product.

[0005] Therefore, developing more novel modification methods for the hydroxyl functional group to design and synthesize a class of novel magnolia phenol derivatives, realizing the development of new anti-myocardial injury drugs, and further applying them to tumor chemotherapy research has important clinical significance and practical value. Summary of the Invention

[0006] Based on this, it is necessary to provide a magnolol derivative and a preparation method and application thereof. The provided magnolol derivative can significantly inhibit H2O2 oxidative damage to myocardial H9C2 cells.

[0007] The first aspect of the present invention provides a magnolol derivative, or a pharmaceutically acceptable salt thereof, having the following structure:

[0008] .

[0009] The second aspect of the present invention provides a method for preparing a magnolol derivative or a pharmaceutically acceptable salt thereof, characterized in that it comprises the following steps:

[0010] Mixing the compound of formula I, the compound of formula II, tri-n-butylphosphine, an azo reagent and a solvent to undergo a nucleophilic substitution reaction to prepare the magnolol derivative or a pharmaceutically acceptable salt thereof;

[0011] The structure of the compound of formula I is shown below:

[0012] ;

[0013] The structure of the compound of formula II is shown below:

[0014] ;

[0015] The structure of the magnolol derivative or a pharmaceutically acceptable salt thereof is shown below:

[0016] .

[0017] In some embodiments, the molar ratio of the compound of formula I to the compound of formula II is (1-2):(1-2). Furthermore, the molar ratio of the compound of formula I to the compound of formula II is 1:(1-2). It is understood that in the present invention, the molar ratio of the compound of formula I to the compound of formula II includes but is not limited to 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, and 1:2.0.

[0018] In some embodiments, the molar ratio of the compound of formula I to the azo reagent and tri-n-butylphosphine is 1:(1.2-2.0):(1.0-1.5). Furthermore, the molar ratio of the compound of formula I to the azo reagent and tri-n-butylphosphine is 1:(1.4-1.8):(1.2-1.4). It can be understood that in the present invention, the molar ratio of the compound of formula I to the azo reagent and tri-n-butylphosphine includes but is not limited to: 1:1.4:1.2; 1:1.4:1.3; 1:1.4:1.4; 1:1.5:1.2; 1:1.5:1.3; 1:1.5:1.4; 1:1.6:1.2; 1:1.6:1.3; 1:1.6:1.4; 1:1.7:1.2; 1:1.7:1.3; 1:1.7:1.4; 1:1.8:1.2; 1:1.8:1.3; 1:1.8:1.4.

[0019] In some embodiments, the nucleophilic substitution reaction time is 20 hours to 30 hours. It is understandable that in the present invention, the nucleophilic substitution reaction time includes but is not limited to 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, and 30 hours.

[0020] In some embodiments, the method for preparing the magnolol derivative or a pharmaceutically acceptable salt thereof is characterized in that the temperature of the nucleophilic substitution reaction is 4°C to 35°C. Furthermore, the temperature of the nucleophilic substitution reaction is 10°C to 30°C. It is understood that in the present invention, the temperature of the nucleophilic substitution reaction includes but is not limited to: 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, and 30°C.

[0021] In some embodiments, the azo reagent is selected from one or more of diethyl azodicarboxylate and diisopropyl azodicarboxylate.

[0022] In some embodiments, the solvent is selected from one or more of tetrahydrofuran and dichloromethane.

[0023] Optionally, the mixing of the compound of formula I, the compound of formula II, tri-n-butylphosphine, an azo reagent and a solvent comprises the following steps:

[0024] mixing the compound of formula I and a portion of the solvent to obtain a first solution;

[0025] mixing the compound of formula II, the azo reagent and the remaining solvent to obtain a second solution;

[0026] The first solution, tri-n-butylphosphine, and the second solution were mixed.

[0027] In some embodiments, the concentration of the compound of formula I in the first solution is 0.1 mmol / mL to 0.9 mmol / mL. Further, the concentration of the compound of formula I in the first solution is 0.1 mmol / mL to 0.6 mmol / mL. It is understood that in the present invention, the concentration of the compound of formula I in the first solution includes but is not limited to 0.1 mmol / mL, 0.2 mmol / mL, 0.3 mmol / mL, 0.4 mmol / mL, 0.5 mmol / mL, and 0.6 mmol / mL.

[0028] In some embodiments, the concentration of the compound of Formula II in the second solution is 0.1 mmol / mL to 0.6 mmol / mL. Furthermore, the concentration of the compound of Formula II in the second solution is 0.1 mmol / mL to 0.4 mmol / mL. It is understood that in the present invention, the concentration of the compound of Formula II in the second solution includes but is not limited to 0.1 mmol / mL, 0.2 mmol / mL, 0.3 mmol / mL, and 0.4 mmol / mL.

[0029] In some embodiments, after the nucleophilic substitution reaction is completed, a post-treatment step is further included, and the post-treatment comprises the following steps: quenching the reaction with dilute hydrochloric acid and water, extracting the resulting reaction solution with dichloromethane, washing the resulting organic phase with water and saturated brine, drying with anhydrous sodium sulfate, filtering, and concentrating after filtration.

[0030] In some embodiments, after the post-treatment, a separation and purification step is further included, and the separation and purification step is: using a silica gel column to separate the intermediate product obtained from the post-treatment to obtain the magnolol derivative or a pharmaceutically acceptable salt thereof.

[0031] The third aspect of the present invention provides a drug for treating myocardial injury, comprising the magnolol derivative as described above, or a pharmaceutically acceptable salt thereof; or comprising the magnolol derivative prepared by the preparation method as described above, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0032] The fourth aspect of the present invention provides a magnolol derivative as described above, or a pharmaceutically acceptable salt thereof; or a magnolol derivative prepared according to the preparation method as described above, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier for use in the preparation of a drug for treating myocardial injury and related diseases.

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] This invention achieves the first chemical splicing of magnolol and idebenone through chemical methods, yielding a novel magnolol derivative, GYS17. Biological evaluation revealed that the magnolol derivative GYS17 significantly inhibits H2O2 oxidative damage in myocardial H9C2 cells, with significantly greater efficacy than the positive controls salvianolate and idebenone. The synthesis of this magnolol derivative has important theoretical and practical implications for the development of novel anti-myocardial injury drugs. This technology also provides new insights and approaches for the structural modification of magnolol derivatives. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1is the magnolol derivative GYS17 in the preparation method of the embodiment of the present invention 1 H NMR spectrum.

[0036] Figure 2 is the magnolol derivative GYS17 in the preparation method of the embodiment of the present invention 13 C NMR spectrum.

[0037] Figure 3 This is the mass spectrum of the magnolol derivative GYS17 in the preparation method of an embodiment of the present invention. DETAILED DESCRIPTION

[0038] The following is a further detailed description of the magnolol derivatives of the present invention, their preparation methods, and applications, with reference to specific examples. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a more thorough and comprehensive understanding of the present disclosure.

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in this specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0040] the term

[0041] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0042] As used herein, "pharmaceutically acceptable" refers to those ligands, materials, compositions, and / or dosage forms that are suitable for administration to a patient within the scope of sound medical judgment and commensurate with a reasonable benefit / risk ratio.

[0043] As used herein, "pharmaceutically acceptable salt" refers to a salt of any compound of the structure shown with an acid or base that is suitable for use as a pharmaceutical. Pharmaceutically acceptable salts include inorganic salts and organic salts. One type of salt is a salt formed by a compound of the present invention with an acid. Suitable acids for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid. Another type of salt is a salt formed by the compound of the present invention and a base. Suitable bases for forming salts include, but are not limited to, alkali metal salts (e.g., sodium salts or potassium salts), alkaline earth metal salts (e.g., magnesium salts or calcium salts), ammonium salts (e.g., lower alkanolammonium salts and other pharmaceutically acceptable amine salts), for example, methylamine salts, ethylamine salts, propylamine salts, dimethylamine salts, trimethylamine salts, diethylamine salts, triethylamine salts, tert-butylamine salts, ethylenediamine salts, hydroxyethylamine salts, dihydroxyethylamine salts, trihydroxyethylamine salts, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0044] As used herein, a "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. As used herein, the language "pharmaceutically acceptable carrier" includes buffers, sterile water for injection, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, that are compatible with drug administration. Each carrier must be "pharmaceutically acceptable" in the sense of being compatible with the other ingredients in the formulation and not harmful to the patient. Suitable examples include, but are not limited to: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch, potato starch, and substituted or unsubstituted β-cyclodextrins; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, cornstarch, and tartar. Rice oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer; (21) other non-toxic compatible substances used in pharmaceutical formulations.

[0045] As used herein, "one or more" refers to any one, any two, or any two or more of the listed items.

[0046] Herein, the optional scope of "and / or", "or / and", and "and / or" includes any one of two or more relevant listed items, and also includes any and all combinations of the relevant listed items, and the said any and all combinations include any two relevant listed items, any more relevant listed items, or a combination of all relevant listed items.

[0047] Herein, “preferred”, “better”, “better”, etc. are only used to describe implementation methods or examples with better effects. It should be understood that they do not constitute limitations on the scope of protection of the present invention.

[0048] Herein, “further”, “further”, “particularly”, etc. are used for descriptive purposes to indicate differences in content, but should not be understood as limiting the scope of protection of the present invention.

[0049] In this document, terms such as "first aspect," "second aspect," "third aspect," and "fourth aspect" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor as implicitly indicating the importance or quantity of the technical features indicated. Furthermore, terms such as "first," "second," "third," and "fourth" serve only as non-exhaustive enumerations and should not constitute closed-ended limitations on quantity.

[0050] In the present invention, when referring to numerical ranges, unless otherwise specified, the numerical ranges are considered continuous and include the minimum and maximum values ​​of the range, as well as every value between such minimum and maximum values. Furthermore, when a range refers to an integer, every integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe a feature or characteristic, the ranges can be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges subsumed therein.

[0051] Unless otherwise specified, the percentage contents mentioned in the present invention refer to mass percentage for solid-liquid mixture and solid-solid mixture, and refer to volume percentage for liquid-liquid mixture.

[0052] The percentage concentrations mentioned in the present invention, unless otherwise specified, refer to the final concentration, which refers to the percentage of the added component in the system after the addition of the component.

[0053] The temperature parameters in the present invention, unless otherwise specified, allow for both constant temperature treatment and treatment within a certain temperature range. The constant temperature treatment allows for temperature fluctuations within the precision range of instrument control. Room temperature in the present invention refers to no temperature control operation, primarily 4°C to 35°C.

[0054] In the present invention, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0055] Using traditional Chinese medicine monomers or their backbones as combinatorial chemistry templates to design and synthesize novel active molecules has proven to be a highly effective drug discovery approach. Magnolol, a biphenyldiphenol compound isolated from the traditional Chinese medicinal herb Magnolia officinalis, exhibits multiple activities, including anti-myocardial injury, anti-tumor, and antioxidant activities. Idebenone, a compound structurally similar to coenzyme Q10, possesses antioxidant activity and plays a protective role on cell membranes and mitochondria, thereby exerting significant cardiovascular protection. Salvianolates exhibit excellent cardiovascular protection and are widely used in the clinical treatment of myocardial injury. Therefore, in this embodiment of the present invention, idebenone and magnolol are used as raw materials. A nucleophilic substitution reaction is used to directly connect the two at the phenolic hydroxyl group, achieving the first chemical splicing of magnolol-idebenone, resulting in a novel magnolol derivative, GYS17. Biological evaluation revealed that the magnolol derivative GYS17 exhibited significant protective effects against H2O2-induced myocardial H9C2 cells, significantly more potent than the positive control, salvianolate.

[0056] The following is further described in conjunction with specific examples. Unless otherwise specified, the raw materials involved in the following specific examples can all be sourced from commercial sources; the instruments used can all be sourced from commercial sources unless otherwise specified; and the processes involved can all be selected conventionally by those skilled in the art unless otherwise specified.

[0057] 1. Preparation of magnolol derivative GYS17

[0058] Example 1

[0059]

[0060] Under argon protection, the compound of formula I (248 mg, 0.93 mmol) was dissolved in dichloromethane (3 mL) and added to the flask, followed by the addition of tri-n-butylphosphine (268 mg, 1.33 mmol). The mixture was stirred in an ice bath and slowly added dropwise with a dichloromethane (5 mL) solution of diisopropyl azodicarboxylate (0.26 ml, 1.4 mmol) and the compound of formula II (314 mg, 0.93 mmol). The mixture was transferred to room temperature and reacted for 27 hours. After the reaction, the mixture was quenched with 1 M dilute hydrochloric acid, quenched with water, extracted with dichloromethane, and the organic phase was washed with water and saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and separated on a silica gel column (petroleum ether / ethyl acetate, 10:1) to obtain an orange-yellow oily liquid GYS17 (462 mg, yield 85%). f= 0.69 (petroleum ether / ethyl acetate, 3:1).

[0061] 1 H NMR (400 MHz, CDCl3) δ 7.16 (d, J = 7.2 Hz, 2H), 7.12 – 7.05 (m,2H), 6.96 (t, J = 8.0 Hz, 2H), 6.59 (s, 1H), 5.98 (dtt, J = 16.5, 9.8, 6.7Hz, 2H), 5.12 – 5.03 (m, 4H), 4.01 (t, J = 6.6 Hz, 2H), 3.98 (d, J = 1.1 Hz, 6H), 3.38 (dd, J = 6.5, 4.9 Hz, 4H), 2.48 – 2.40 (m, 2H), 2.01 (s, 3H), 1.75– 1.69 (m, 2H), 1.42 – 1.35 (m, 2H), 1.30 (d, J = 13.5 Hz, 6H), 1.26 (s, 3H), 1.22 (s, 3H).

[0062] 13 C NMR (100 MHz, CDCl3) δ 184.67, 184.10, 153.21, 152.17, 144.24,144.22, 143.03, 138.61, 137.86, 137.41, 133.90, 132.55, 132.17, 131.17,129.14, 128.95, 127.91, 126.46, 117.66, 115.78, 115.41, 113.48, 70.02, 61.09,39.40, 39.33, 29.75, 29.32, 29.28, 29.26, 29.09, 28.97, 28.67, 26.34, 25.66,11.86.

[0063] MS (ESI) m / z = 609.1 [M+Na] + .

[0064] Example 2

[0065] This example provides a method for preparing a magnolol derivative GYS17. The steps are the same as those in Example 1, with the main difference being that diethyl azodicarboxylate is used in equimolar substitution for diisopropyl azodicarboxylate.

[0066] Results: The magnolol derivative GYS17 could be prepared by replacing diisopropyl azodicarboxylate with diethyl azodicarboxylate in equal moles, and the yield was reduced to 70%.

[0067] Example 3

[0068] This example provides a preparation method for a magnolol derivative GYS17. The steps are the same as those in Example 1, with the main difference being that an equal volume of tetrahydrofuran is used instead of dichloromethane.

[0069] Results: The magnolol derivative GYS17 could be prepared by replacing dichloromethane with an equal volume of tetrahydrofuran, with a yield reduced to 40%.

[0070] Comparative Example 1

[0071] This comparative example provides a preparation method of a magnolol derivative GYS17, and its synthesis steps are as follows:

[0072] Under argon protection, the compound of formula II (338 mg, 1 mmol) was dissolved in dichloromethane (5 mL), and p-toluenesulfonyl chloride (190 mg, 1 mmol), triethylamine (0.14 mL, 1 mmol), 25 o C for 24 hours, and a tetrahydrofuran (2 mL) solution of the compound of formula I (266 mg, 1 mmol) was slowly added. After 24 hours of reaction, thin-layer chromatography monitoring revealed that the reaction system was disordered, and the reaction was quenched with water. The mixture was extracted with dichloromethane, and the organic phase was washed with water and saturated brine. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column, but the magnolol derivative GYS17 could not be obtained.

[0073] Comparative Example 2

[0074] This comparative example provides a preparation method for a magnolol derivative GYS17. The synthesis steps are the same as those in comparative example 1, with the main difference being that methanesulfonyl chloride is used in equimolar replacement of p-toluenesulfonyl chloride. Thin layer chromatography monitoring revealed that the reaction system was disordered, and as a result, the magnolol derivative GYS17 could not be obtained.

[0075] The reaction condition screening results of the embodiments and comparative examples provided by the present invention are shown in Table 1.

[0076] Table 1 Reaction condition screening results

[0077]

[0078] 2. Biological Evaluation

[0079] (I) Effects of H2O2 on the cell viability of rat cardiomyocytes H9C2 cell line

[0080] 1. Experimental Materials

[0081] 1.1 Cell line: Rat cardiomyocyte H9C2 cell line.

[0082] 1.2 Reagents and instruments: H2O2 solution (Guangdong Hengjian Pharmaceutical Co., Ltd.); 0.25% trypsin (GIBCO); DMEM medium (Beijing Solebold Technology Co., Ltd.); fetal bovine serum (GIBCO); CCK8 kit (ZETA); dimethyl sulfoxide (DMSO, Beijing Solebold Technology Co., Ltd.); low-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.); constant temperature water bath (Shanghai Jinghong Laboratory Equipment Co., Ltd.); clean bench (Singapore Yisi High-Tech Co., Ltd.); cell culture incubator (Singapore Yisi High-Tech Co., Ltd.); inverted microscope (Leica); HERAcell vios 250i cell constant temperature incubator (Thermo Fisher Scientific, USA); imark microplate reader (Bio-Rad, USA).

[0083] 2. Experimental Methods

[0084] 2.1 Culture H9C2 cells. When they reach the logarithmic growth phase, digest them with 0.25% trypsin and centrifuge at 900 rpm for 3 min. Discard the supernatant and resuspend the cell pellet in DMEM medium containing 10% FBS. Plate the cells at a density of 5000 cells / well in a 96-well culture plate and place them in a 37 o C, 5% CO2 and saturated humidity for 24 h.

[0085] 2.2 Set up 6 replicate wells in each group, add 100 μL of DMEM medium and H2O2 solution to each well according to the following conditions, and then continue culturing.

[0086] Control group: the group without adding H2O2 solution.

[0087] Model group 1: 2 μL of 0.1 mol / L H2O2 solution was added.

[0088] Model group 2: 3 μL of 0.1 mol / L H2O2 solution was added.

[0089] Model group 3: 4 μL of 0.1 mol / L H2O2 solution was added.

[0090] Model group 4: 5 μL of 0.1 mol / L H2O2 solution was added.

[0091] Model group 5: 6 μL of 0.1 mol / L H2O2 solution was added.

[0092] Model group 6: 7 μL of 0.1 mol / L H2O2 solution was added.

[0093] After 24 hours of culture, the effects of H2O2 on H9C2 cells were assessed using a CCK-8 assay kit. The procedure was as follows: 10 μL of CCK8 reagent was added to each well, and the cells were cultured for another 2 hours. The absorbance (OD) was measured at 450 nm using a microplate reader. The cell survival rate of each model group was calculated, assuming the cell survival rate of the control group was 100%.

[0094] 3. Experimental Results

[0095] Table 2 Effects of H2O2 on the cell viability of rat cardiomyocytes H9C2 cell line

[0096]

[0097] The experimental results are shown in Table 2. H2O2 solution can significantly oxidatively damage rat myocardial H9C2 cells, and the damage effect increases with increasing dose. When the dose of 0.1 mol / L H2O2 solution is 5 μL, the survival rate of rat myocardial H9C2 cells decreases to 50.74%. This dose can be used to construct an H9C2 cell oxidative damage model.

[0098] (II) Effects of magnolol derivative GYS17 on myocardial H9C2 cells damaged by H2O2

[0099] 1. Experimental Materials

[0100] (1) Cell line: rat cardiomyocyte H9C2 cell line.

[0101] (2) Reagents and instruments: H2O2 solution (Guangdong Hengjian Pharmaceutical Co., Ltd.); 0.25% trypsin (GIBCO); DMEM culture medium (Beijing Solebao Technology Co., Ltd.); fetal bovine serum (GIBCO); CCK8 kit (ZETA); dimethyl sulfoxide (DMSO, Beijing Solebao Technology Co., Ltd.); low-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.); constant temperature water bath (Shanghai Jinghong Laboratory Equipment Co., Ltd.), clean bench (Singapore Yisi High-Tech Co., Ltd.); cell culture incubator (Singapore Yisi High-Tech Co., Ltd.), inverted microscope (Leica); HERAcell vios 250i cell constant temperature incubator (Thermo Fisher Scientific, USA); imark microplate reader (Bio-Rad, USA).

[0102] 2. Experimental Methods

[0103] 2.1 Culture H9C2 cells until they reach the logarithmic growth phase, digest them with 0.25% trypsin, centrifuge at 900 rpm for 3 min, discard the supernatant, resuspend the cell pellet in DMEM medium containing 10% FBS, and seed the cells at a density of 5000 cells / well in a 96-well culture plate. o C, 5% CO2 and saturated humidity for 24 h.

[0104] 2.2 A control group, model group, positive group, idebenone group, and GYS17 group were set up. Six replicate wells were set up in each group. After incubation for 5 h with drug addition according to the following conditions, the original culture medium was removed and 100 μL of DMEM medium was added. Furthermore, 5 μL of 0.1 mol / L H2O2 solution was added to damage the cells, and the cells were incubated for 24 h.

[0105] Control group: no medication, no injury group.

[0106] Model group: After incubation for 5 h without drug administration, 5 μL of 0.1 mol / L H2O2 solution was added to damage the cells.

[0107] Positive group: 5 μM salvianolate was added, and after incubation for 5 h, the original culture medium was removed and 100 μL DMEM culture medium was added, followed by 5 μL of 0.1 mol / L H2O2 solution to damage the cells.

[0108] Idebenone group: 5 μM idebenone was added, and after incubation for 5 h, the original culture medium was removed and 100 μL DMEM culture medium was added, followed by 5 μL of 0.1 mol / L H2O2 solution to damage the cells.

[0109] The GYS17 treatment group was divided into 3 groups:

[0110] GYS17 group 1: 1 μM GYS17 was added, and after incubation for 5 h, the original culture medium was removed and 100 μL DMEM culture medium was added, followed by 5 μL of 0.1 mol / L H2O2 solution to damage the cells.

[0111] GYS17 group 2: 5 μM GYS17 was added, and after incubation for 5 h, the original culture medium was removed and 100 μL DMEM culture medium was added, followed by 5 μL of 0.1 mol / L H2O2 solution to damage the cells.

[0112] GYS17 group 3: 10 μM GYS17 was added, and after incubation for 5 h, the original culture medium was removed and 100 μL DMEM culture medium was added, followed by 5 μL of 0.1 mol / L H2O2 solution to damage the cells.

[0113] After 24 hours of cell culture, the effects of the drugs on the cells were detected using a CCK-8 kit. The procedure was as follows: 10 μL of CCK-8 reagent was added to each well, and the cells were cultured for another 2 hours. The absorbance (OD) was measured at a wavelength of 450 nm using a microplate reader. The cell viability of the control group was set as 100%, and the cell viability of the other groups was calculated.

[0114] 3. Experimental Results

[0115] Table 3 Protective effect of magnolol derivative GYS17 on myocardial H9C2 cells oxidatively damaged by H2O2

[0116]

[0117] The experimental results are shown in Table 3. The magnolol derivative GYS17 provided by the present invention can significantly inhibit the oxidative damage of myocardial H9C2 cells, and the effect is significantly better than the positive control drug salvianolate and the prototype drug idebenone.

[0118] The present invention relates to a magnolol derivative and a preparation method and application thereof. The magnolol derivative comprises: C 37 H 46 O6 (GYS17). This invention achieves the first chemical splicing of magnolol and idebenone through chemical methods, yielding a novel magnolol derivative, GYS17. Biological evaluation revealed that the magnolol derivative GYS17 significantly inhibits H2O2 oxidative damage in myocardial H9C2 cells, with significantly greater efficacy than the positive controls salvianolate and idebenone. The synthesis of this magnolol derivative has important theoretical and practical implications for the development of novel anti-myocardial injury drugs. This technology also provides new insights and approaches for the structural modification of magnolol derivatives.

[0119] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0120] The above-described embodiments merely represent several implementation methods of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art may make various modifications and improvements without departing from the spirit of the present invention, and these modifications and improvements fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be based on the appended claims, and the description may be used to interpret the content of the claims.

Claims

1. A magnolol derivative, or a pharmaceutically acceptable salt thereof, characterized in that: The magnolol derivative has the following structure: 。 2. A method for preparing a magnolol derivative, characterized in that: The following steps are involved: Mixing the compound of formula I, the compound of formula II, tri-n-butylphosphine, an azo reagent and a solvent to undergo a nucleophilic substitution reaction to prepare the magnolol derivative; The structure of the compound of formula I is shown below: ； The structure of the compound of formula II is shown below: ； The structure of the magnolol derivative is shown below: ； The azo reagent is selected from one or more of diethyl azodicarboxylate and diisopropyl azodicarboxylate; The solvent is selected from one or more of tetrahydrofuran and dichloromethane.

3. The method for preparing a magnolol derivative according to claim 2, wherein The molar ratio of the compound of formula I to the compound of formula II is (1-2): (1-2).

4. The method for preparing a magnolol derivative according to claim 2, wherein The molar ratio of the compound of formula I to the azo reagent and the tri-n-butylphosphine is 1:(1.2-2.0):(1.0-1.5).

5. The method for preparing a magnolol derivative according to claim 2, wherein The time of the nucleophilic substitution reaction is 20 hours to 30 hours.

6. The method for preparing a magnolol derivative according to claim 2, wherein The temperature of the nucleophilic substitution reaction is 4°C to 35°C.

7. A drug for treating myocardial oxidative damage, characterized in that: The invention comprises the magnolol derivative according to claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

8. Use of the magnolol derivative according to claim 1, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating myocardial oxidative damage.

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