Preparation and application of novel magnolol and jateorhizine conjugate

Abstract

The invention discloses preparation and application of a novel magnolol and jatrorrhizine conjugate, relates to the technical field of medicines, and aims to overcome the defects of poor drug resistance and insufficient activity of a single-target anti-cancer drug. According to the technical scheme, acetone or acetonitrile is used as a solvent, and through a two-step substitution reaction, the magnolol and jatrorrhizine conjugate is obtained; the preparation method comprises the following steps: reacting magnolol with dibromo-alkane to generate an intermediate B, and conjugating the intermediate B with jatrorrhizine to obtain a final product with a general formula C; or the jatrorrhizine reacts with dibromo alkane to generate an intermediate E, and then the intermediate E is conjugated with magnolol to obtain the final product of the general formula C. In the preparation process, reaction conditions (such as potassium carbonate alkaline environment and 82 DEG C reflux) are optimized, and high yield and purity are realized. The conjugate disclosed by the invention is widely applied to the anti-tumor field, has remarkable inhibitory activity on various cancer cells (such as non-small cell lung cancer cells HCC827 and human lung adenocarcinoma cells H1975), has better effects than magnolol and jateorhizine monomers, and provides a new candidate for research and development of multi-target drugs.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the preparation and application of a novel magnolol and herb root alkaloid conjugate. Background Technology

[0002] Cancer is a common clinical disease caused by abnormal cell growth leading to heterogeneity in the body. Cancer is one of the most important diseases threatening human life and health today. A report released by the Global Cancer Institute shows that the number of people who die from cancer in Asia accounts for about 55% of the total number of cancer deaths. In recent years, a large number of active natural components from plants have become an important way to discover innovative anti-tumor drugs. Current research shows that more than 60 percent of drug small molecules can be traced back to the inspiration of natural products or the natural products themselves. Multi-target anticancer drugs can improve the therapeutic effect of single-target anticancer drugs and overcome the resistance of single-target inhibitors. It is an important research direction for the development of anticancer drugs. Jatrorrhizine is also a quaternary ammonium isoquinoline alkaloid with multiple activities such as antifungal, hypoglycemic, and antitumor [1]. Magnoliae officinalis is one of the main components of the traditional Chinese medicine Magnolia officinalis. Magnoliae officinalis has antioxidant, antidepressant, antibacterial and antitumor effects.

[0003] Due to the complexity of tumor pathogenesis and the problem of drug resistance in targeted therapies, single-target drugs are no longer effective in treating cancer. Natural products, with their advantages of structural diversity, high activity, and low toxicity, have become an important source for the discovery of multi-target drugs. Conjugate design is an innovative synthetic strategy; combining natural products can allow different pharmacophores to act on different targets, enhancing the antitumor activity of natural products and representing a crucial pathway and strategy for discovering innovative drugs.

[0004] Therefore, we propose a method for the preparation and application of a compound of purslane and magnolol. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to overcome the existing defects and provide a new preparation and application of magnolol and purslane conjugates, which can form multi-target conjugates by combining magnolol and purslane to synergistically enhance antitumor activity, and can effectively solve the problems in the background art.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a novel magnolol and herb root alkaloid conjugate, comprising the following general structural formula I:

[0007]

[0008] Wherein: R is magnolol or a derivative of magnolol, n is a natural number greater than 1, defining the core structure of the conjugate, achieving multi-target anti-tumor effects by combining magnolol and purslane, improving activity and overcoming drug resistance.

[0009] Furthermore, n is an integer from 2 to 10, balancing the linker chain length to optimize bioactivity and synthetic feasibility, and avoiding steric hindrance.

[0010] A novel method for preparing a magnolol and herb root alkaloid conjugate includes the following steps:

[0011]

[0012] (a) Using acetone or acetonitrile as solvent, magnolol and dibromoalkanes as reactants in a molar ratio of 1:1 to 1.5, react in the presence of potassium carbonate at 60°C or 82°C for 6-12 hours to obtain intermediate B.

[0013] (b) Using acetonitrile, tetrahydrofuran or N,N-dimethylformamide as solvent, intermediate B and purslane as reactants in a molar ratio of 1:1 to 1.5, the reaction is carried out under reflux at 82°C in the presence of potassium carbonate to obtain the final product of general formula C.

[0014] (c) Using acetonitrile as solvent, purslane and dibromoalkanes as reactants in a molar ratio of 1:1 to 1.2, the reaction is carried out under reflux at 82°C in the presence of potassium carbonate to prepare intermediate E.

[0015] (d) Using acetonitrile as solvent, intermediate E and magnolol as reactants in a molar ratio of 1:1, the reaction was carried out under reflux at 82°C in the presence of potassium carbonate to obtain the final product of general formula C.

[0016] Furthermore, in step (a), the molar ratio of magnolol, dibromoalkanes and potassium carbonate is 1:1.2:1.8, the reaction temperature is 82°C, and the reaction time is 8 hours. These refined conditions improve the yield and ensure that the reaction is complete and of high purity.

[0017] Furthermore, in step (b), the molar ratio of intermediate B, purslane, and potassium carbonate is 1.2:1:1.5, the solvent is acetonitrile, and the reflux time is 6 hours. The excess of intermediate B (1.2:1) drives the reaction, and the acetonitrile solvent and 6-hour reflux ensure efficient synthesis.

[0018] Furthermore, in step (c), the molar ratio of purslane, 1,10-dibromodecane, and potassium carbonate is 1:1.2:1.5, the solvent is acetonitrile, and the reflux time is 6 hours, which aims to improve the efficiency and yield of purslane modification along the purslane modification pathway.

[0019] Furthermore, in step (d), the molar ratio of intermediate E, magnolol, and potassium carbonate is 1:1:1.2, the solvent is acetonitrile, and the reflux time is 6 hours to ensure the purity of the final product C.

[0020] A novel magnolol and magnolol alkaloid conjugate or a novel method for preparing a novel magnolol and magnolol alkaloid conjugate, and the application of the novel magnolol and magnolol alkaloid conjugate in the field of anti-tumor therapy, including human liver cancer cells MHCC97H, human lung adenocarcinoma cells H1975, non-small cell lung cancer cells HCC827, and cervical cancer cells HELA.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows: The preparation and application of the novel magnolol and herb root alkaloid conjugate have the following advantages:

[0022] Utilizing the principles of drug design and synthesis, a series of novel rutin-honokiol conjugates were designed and synthesized for the first time by hybridizing the antitumor-active natural product, rutin, with magnolol. By controlling the molar ratio of reactants using dibromoalkanes of varying carbon chain lengths, a series of rutin-honokiol conjugates were synthesized via a two-step substitution reaction. The resulting compounds exhibited significantly superior inhibitory activity against tumor cell proliferation compared to the monomeric lead compounds of rutin and magnolol, providing a more potent candidate drug for antitumor drug development. Detailed Implementation

[0023] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0024] This invention provides a technical solution: a novel magnolol and herb root alkaloid conjugate, comprising the following general structural formula I:

[0025]

[0026] Wherein: R is magnolol or a derivative of magnolol, n is a natural number greater than 1, n is an integer from 2 to 10, the chain length is balanced to optimize biological activity and synthetic feasibility, avoid steric hindrance, define the core structure of the conjugate, achieve multi-target anti-tumor effects by combining magnolol and purslane, improve activity and overcome drug resistance.

[0027] A novel method for preparing a magnolol and herb root alkaloid conjugate includes the following steps:

[0028]

[0029] (a) Using acetone or acetonitrile as solvent, magnolol and dibromoalkanes as reactants in a molar ratio of 1:1 to 1.5, reacting at 60°C or 82°C for 6-12 hours in the presence of potassium carbonate, intermediate B is obtained. The molar ratio of magnolol, dibromoalkanes and potassium carbonate is 1:1.2:1.8, the reaction temperature is 82°C and the reaction time is 8 hours. Refining the conditions improves the yield and ensures complete reaction and high purity.

[0030] (b) Using acetonitrile, tetrahydrofuran or N,N-dimethylformamide as solvent, intermediate B and purslane as reactants in a molar ratio of 1:1 to 1.5, the reaction is carried out under reflux at 82°C in the presence of potassium carbonate to obtain the final product of general formula C. The molar ratio of intermediate B, purslane and potassium carbonate is 1.2:1:1.5, the solvent is acetonitrile, and the reflux time is 6 hours. The reaction is driven by excess intermediate B (1.2:1), and the acetonitrile solvent and 6 hours of reflux ensure efficient synthesis.

[0031] (c) Using acetonitrile as solvent, purslane and dibromoalkane as reactants in a molar ratio of 1:1 to 1.2, the reaction was carried out under reflux at 82°C in the presence of potassium carbonate to prepare intermediate E. The molar ratio of purslane, 1,10-dibromodecane and potassium carbonate was 1:1.2:1.5, the solvent was acetonitrile, and the reflux time was 6 hours. This method is aimed at improving the efficiency and yield of purslane modification along the purslane modification pathway.

[0032] (d) Using acetonitrile as solvent, intermediate E and magnolol as reactants in a molar ratio of 1:1, the reaction was carried out under reflux at 82°C in the presence of potassium carbonate to obtain the final product of general formula F. The molar ratio of intermediate E, magnolol and potassium carbonate was 1:1:1.2, the solvent was acetonitrile, and the reflux time was 6 hours to ensure the purity of the final product F.

[0033] This invention discloses the application of a novel magnolol and magnolol alkaloid conjugate prepared by a novel method, and its application in the field of antitumor therapy, including tumors such as human liver cancer cells MHCC97H, human lung adenocarcinoma cell line H1975, non-small cell lung cancer cells HCC827, and cervical cancer cells HELA. The invention provides the following examples:

[0034] Example 1: Target compound C1

[0035]

[0036] Preparation process of target compound C1:

[0037]

[0038] (a) Preparation of intermediate B1

[0039] Weigh out magnolol [C 18 H 18 [O2] (1.2 g, 4.5 mmol) was placed in a 50 mL round-bottom flask, 25 mL of acetone was added, followed by 0.32 g of anhydrous K2CO3. The mixture was stirred at room temperature for 2 h, and the reaction was stopped. Then, 0.467 mL of 1,2-dibromoethane (ρ = 2.18 g / cm3) was added, and the mixture was refluxed at 40 °C for 20 h. After stopping the reaction and cooling, potassium carbonate was removed, and the product was concentrated to obtain the crude product. The crude product was then subjected to silica gel column chromatography with petroleum ether:ethyl acetate (30:1) as elution to give a yellow oily intermediate 16. [C] 20 H 21 BrO2]ESIMS: m / z 395.0, [M+Na] + ; 1 HNMR(600MHz, CDCl3)δ:7.22-7.19(m,2H,Ben-H),7.15(dd,J=8.2,2.2Hz,1H,Ben-H),7 .10(d,J=2.2Hz,1H,Ben-H),7.01(t,J=8.1Hz,2H,Ben-H),6.07-6.01(m,3H,-CH-C),5.1 4-5.12(m,2H,-CH2=C),5.10-5.08(m,2H,-CH2Br),4.30(t,J=6.3Hz,2H,-OCH2-),3.54 (t,J=6.3Hz,2H,-CH2Br), 3.43(d,J=6.7Hz,2H,PhCH2-), 3.40(d,J=6.7Hz,2H,PhCH2-).

[0040] (ii) Preparation of final product C1

[0041] Weigh out intermediate 17[C] 20 H 21 BrO2] (100.0 mg, 0.269 mmol), purslane [C 20 H 20 NO4]+ (82.66 mg, 0.245 mmol) was added to a 100 mL round-bottom flask, along with the alkaloid and 50 mL acetonitrile. After complete dissolution, 0.26 g of anhydrous K2CO3 was added, and the mixture was refluxed at 83 °C for 6 h. Heating was then stopped, and the reaction was allowed to cool. Potassium carbonate was filtered off, and the mixture was concentrated to obtain the crude product. The crude product was then subjected to 300-400 mesh Al2O3 column chromatography with CH2Cl2:CH3OH (30:1) as elution. Thin-layer chromatography was performed under the following conditions: toluene:ethyl acetate:isopropanol:methanol:ammonia (6:3:3:1.5:0.5) to give a yellow solid final product. Cl.Mp: 151.3 °C~152.3 °C. ESIMS (m / z) 630.3 [M-Cl]+ ; 1 HNMR(600MHz,DMSO-d6)δ:9.89(s,1H,H-8″′),9.04(s,1H,H-13″′),9.01(s,-OH,1H,H-6′),8.21(d,J=9.1Hz,1H,H-12″′),8.03(d,J=9.1Hz,1H,H-11″′),7.72(s,1H,H-1″′),7.12(d,J=1.3Hz,2H,H-3″,H-4″),7.04(s,2H,H-4′,H-5′),6.96(d,J=2.2Hz,1H,H-4″′),6.89(s,1H,H-6″),6.80(s,1H,H-2′),5.95-5.86(m,2H,H-8′,H-8″),5.12-5.05(m,2H,H-9′),5.00-4.97(m,2H,H-9″),4.92(t,J=6.0Hz,2H,H-6″′),4.34(t,J=2.6Hz,2H,H-1),4.29(t,J=2.6Hz,2H,H-2),4.11(s,-OCH3,3H,H-16″′),4.08(s,-OCH3,3H,H-17″′),3.91(s,-OCH3,3H,H-15″′),3.34(t,J=7.0Hz,2H,H-5″′),

[0042] 3.20(m,4H,H-7′,H-7″); 13CNMR(100MHz,DMSO-d6)δ:154.68(C-2″),153.26(C-6′),151.09(C-10″′),150.76 (C-3″′),149.29(C-14″′),145.98(C-9″′),144.34(C-8″′),144.12(C-2″′),138.8 1(C-12a),138.52(C-8′),138.09(C-8″),133.54(C-3′),132.14(C-5″),131.90(C -2′),129.80(C-13″′),129.04(C-6″),128.53(C-4a),128.47(C-4′),128.42(C-4″ ),127.26(C-1″),125.49(C-1′),123.91(C-12″′),121.84(C-14a),120.38(C-8a) ,119.68(C-5′),116.01(C-3″),115.53(overlap)(C-9″,C-9′),113.99(C-1″′),11 2.83(C-4″′),109.59(C-11″′),68.01(C-1),67.68(C-2),62.38(C-16″′),57.51(C -6″′), 56.7(C-17″′), 55.84(C-15″′), 39.17(C-7′), 39.14(C-7″), 26.40(C-5″′).

[0043] Example 2: Target compound C2

[0044]

[0045] Preparation process of target compound C2

[0046]

[0047] (b) Preparation of intermediate B2

[0048] Following the method used to prepare intermediate B1, a yellow, oily product B2 was obtained. [C] 24 H 29 BrO2]ESIMS: m / z 451.1 [M+Na] + ; 1HNMR (600MHz, CDCl3) δ: 7.23 (m, 2H, Ben-H), 7.14 (dd, J = 8.2, 2.2Hz, 1H, Ben-H), 7.12 (d ,J=2.2Hz,1H,Ben-H),7.01(t,J=8.1Hz,2H,Ben-H),6.05-6.03(m,2H,-CH2=C),5.17-5. 14(m,4H,-(CH2)-),4.08(t,J=6.3Hz,2H,-OCH2-),3.45(t,J=6.3Hz,4H,-(CH2)2-),3.3 7(d,J=6.7Hz,2H,-CH2Br-),1.85-1.77(m,4H,-(CH2)2-),1.41-1.44(m,4H,-(CH2)2-).

[0049] (b) Preparation of final product C2

[0050] Weigh out intermediate b2[C] separately 24 H 29 BrO2] (102.0 mg, 0.238 mmol) and purslane [C 20 H 20 NO4]+ (85 mg, 0.251 mmol) was added to a 100 mL round-bottom flask, along with the herbicide and 50 mL acetonitrile. After complete dissolution, 0.26 g of anhydrous K2CO3 was added, and the mixture was refluxed at 83 °C for 6 h. Heating was then stopped, and the reaction was allowed to cool. Potassium carbonate was filtered off, and the mixture was concentrated to obtain the crude product. The crude product was then obtained by 300-400 mesh Al2O3 column chromatography with CH2Cl2:CH3OH (30:1) as elution. The thin-layer chromatography conditions were toluene:ethyl acetate:isopropanol:methanol:ammonia (6:3:3:1.5:0.5) to give the final product C2 as a yellow solid with a melting point of 143.0 °C–144.0 °C. [C 44 H 48 NO6] + ESIMS: m / z 686.3 [M-Cl] + ; 1HNMR(600MHz,DMSO-d6)δ:9.89(s,1H,H-8″′),9.06(s,1H,H-13″′),9.01(s,Ben-OH,1H,H-6′,),8.22(d,J=9.1Hz,1H,H-12″′),8.06(d,J=9.1Hz,1H,H-11″′),7.72(s,1H,H-1″′),7.08(d,J=1.3Hz,2H,H-3″,H-4″),6.97(s,2H,H-4′,H-5′),6.94(d,J=2.2Hz,1H,H-4″′),6.91(s,1H,H-6″),6.77(s,1H,H-2′),5.95-5.86(m,2H,H-8′,H-8″),5.10-5.05(m,2H,H-9′),5.03-5.00(m,2H,H-9″),4.94(t,J=6.0Hz,2H,H-6″′),4.34(s,-OCH3,3H,H-16″′),4.29(s,-OCH3,3H,H-17″′),4.11(t,J=2.6Hz,2H,H-1),4.08(s,-OCH3,3H,H-15″′),3.91(t,J=2.6Hz,2H,H-6),3.23(m,4H,H-7′,H-7″),3.21(t,J=7.0Hz,2H,H-5″′),1.79(m,4H,H-2,H-5),1.38-1.59(m,4H,H-3,H-4);

[0051] 13CNMR(100MHz,DMSO-d6)δ:154.89(C-2″),153.34(C-6′),151.41(C-10″′),150.70(C-3″′ ),149.32(C-14″′),145.90(C-9″′),144.09(C-8″′),138.77(C-2″′),138.57(C-12a),13 8.20(C-8′),133.59(C-8″),131.90(C-3′),131.86(C-5″),131.41(C-2′),129.55(C-13″ ),129.06(C-6″),128.42(C-4a),128.36(C-4′),127.25(C-4″),125.71(C-1″),123.89(C -1′),121.81(C-12″′),120.30(C-14a),119.25(C-8a),115.92(C-5′),115.84(C-3″),11 5.65(overlap)(C-9″,C-9′),113.14(C-1″′),112.54(C-4″′),109.39(C-11″′),68.81(C -1),68.32(C-6),62.38(C-16″′),57.52(C-6″′),56.68(C-17″′),55.86(C-15″′),39.21 (C-7′), 39.17(C-7″), 29.13(C-2), 28.97(C-5), 26.43(C-5″′), 25.63(C-3), 25.57(C-4).

[0052] Implementation Example 3 Target Compound F

[0053]

[0054] Preparation process of target compound F

[0055]

[0056] (c) Preparation of intermediate E (n=10) of purslane derivative

[0057]

[0058] Weigh out the root alkaloids [C] 20 H 20 NO4] +(1.6 g, 4.73 mmol) was placed in a 50 mL round-bottom flask, 25 mL of acetonitrile was added, and 0.34 g of anhydrous K₂CO₃ was added. The mixture was stirred at room temperature for 2 h, and then 1.62 mL of 1,10-dibromodecane (ρ = 1.335 g / cm³) was added. The mixture was refluxed at 83 °C for 6 h. The reaction was then stopped, and the mixture was cooled to room temperature. The mixture was then concentrated by suction filtration to obtain the crude product. The product was then subjected to Al₂O₃ column chromatography (30-400 mesh) with CH₂Cl₂:CH₃OH (30:1) as elution. The thin-layer chromatography conditions were toluene:ethyl acetate:isopropanol:methanol:ammonia (6:3:3:1.5:0.5), yielding a yellow crystalline product 22. Mp: 188.8 °C~189.8 °C. ESIMS: m / z 556.2 [M-Cl] + . 1 HNMR(600MHz,DMSO-d6)δ:9.89(s,1H,H-8″′),9.08(s,1H,H-13″′),8.21(s,1H,H-1″′),8.07(d,J=2.2Hz,1H,H-4″′ ),7.71(d,J=9.1Hz,1H,H-12″′),7.09(d,J=9.1Hz,1H,H-11″′),4.97-4.95(t,J=6.3Hz,2H,H-6″′),4.11(s,-OCH3,3 H,H-16″′),4.08(s,-OCH3,3H,H-17″′),4.07(t,J=16.7Hz,2H,H-1),4.06-4.05(s,-OCH3,3H,H-15″′),3.64-3.52( t,J=6.7Hz,2H,H-10),3.23-3.21(t,J=6.3Hz,2H,H-5″′),1.43-1.29(m,16H,H-2,H-3,H-4,H-5,H-6,H-7,H-8,H-9); 13CNMR(100MHz,DMSO-d6)δ:151.40(C-10″′),150.68(C-3″′),49.28(C-14a),145.86(C-9″′),144.06(C-8″′),138.16(C-2″ ′),133.59(C-12a),129.04(C-13″′),127.20(C-4a),123.89(C-12″′),121.79(C-14″′),120.31(C-8a),119.23(C-1″′),11 2.55(C-4″′),109.37(C-111″′),68.90(C-1),62.38(C-16″′),57.51(C-6″′),56.73(C-17″′),55.86(C-15″′),45.91(C-1 0),35.71(C-9),32.71(C-2),29.36(C-4),29.30(C-5),29.14(C-6),29.02(C-5″′),28.56(C-7),27.99(C-8),25.93(C-3).

[0059] (d) Preparation of final product F: Weigh intermediate E[C 30 H 39 BrNO4]+ (190.0 mg, 0.342 mmol), magnolol [C 18 H 18 [O2] (90.0 mg, 3.38 mmol), dissolved completely in 25 mL of acetonitrile, then 0.32 g of anhydrous K2CO3 was added, and the mixture was refluxed at 83 °C for 6 h. Potassium carbonate was filtered off, and the crude product was concentrated. The product was then subjected to Al2O3 column chromatography (300-400 mesh). Elution was performed using CH2Cl2:CH3OH (50:1) under the following conditions: toluene:ethyl acetate:isopropanol:methanol:ammonia (6:3:3:1.5:0.5). A yellow solid final product, 23, was obtained, with a pH of 128.8 °C–129.8 °C. ESIMS (m / z) 742.4 [M-Cl] + . 1HNMR(600MHz,DMSO-d6)δ:9.88(s,1H,H-8″′),9.02(s,1H,H-13″′),9.01(s,Ben-OH,1H,H-6′,),8.21(d,J=9.1Hz,1H,H-12″′),8.03(d,J=9.1Hz,1H,H-11″′),7.70(s,1H,H-1″′),7.08(d,J=1.3Hz,2H,H-3″,H-4″),6.99(s,2H,H-4′,H-5′),6.94(d,J=2.2Hz,1H,H-4″′),6.89(s,1H,H-6″),6.61(s,1H,H-2′),5.94(m,2H,H-8′,H-8″),5.06(m,2H,H-9′),5.02(m,2H,H-9″),4.98(t,J=6.0Hz,2H,H-6″′),4.10(s,-OCH3,3H,H-16″′),4.07(s,-OCH3,3H,H-17″′),3.93(t,J=2.6Hz,2H,H-1),3.86(s,-OCH3,3H,H-15″′),3.79(t,J=2.6Hz,2H,H-10),3.26(m,4H,H-7′,H-7″),3.20(t,J=7.0Hz,2H,H-5″′),1.77-1.74(m,2H,H-2),1.55-1.53(m,2H,H-9),1.39(m,4H,H-3,H-8),1.23(m,8H,H-4,H-5,H-6,H-7); 13 CNMR(100MHz,DMSO-d6)δ:

[0060] 154.84(C-2″),153.38(C-6′),151.41(C-10″′),150.68(C-3″′),149.28(C-14″′),

[0061] 145.88(C-9″′),144.06(C-8″′),139.30(C-2″′),138.78(C-12a),138.57(C-8′),

[0062] 133.56(C-8″),131.86(C-3′),

[0063] 131.29(C-5″),130.74(C-2′),129.40(C-13″),129.06(C-6″),128.37(C-4a),127.94(C-4′),127.19(C-4″),125.61(C-1″),123.91(C- 1′),121.79(C-12″′),120.28(C-14a),119.21(C-8a),117.83(C-5′),115.91(C-3″),115.59(C-9″),115.24C-9′),113.09(C-1″′),112 .49(C-4″′),109.30(C-11″′),68.86(C-1),68.36(C-10),62.36(C-16″′),57.48(C-6″′),56.62(C-17″′),55.85(C-15″′),39.25(C-7′ ),39.18(C-7″),29.42(C-2),29.21(C-4),29.04(C-5),28.22(C-6),26.74(C-7),26.41(C-9),25.94(C-5″′),22.87(C-3),22.57(C-8).

[0064] The method for detecting antitumor activity in this experiment is as follows:

[0065] Cell culture: Human liver cancer cells MHCC97H, human lung adenocarcinoma cell line H1975, non-small cell lung cancer cells HCC827, and cervical cancer cells HELA were cultured in RPMI-1640 and DMEM medium containing 10% FBS (fetal bovine serum) and continuously in an incubator with a temperature of 37℃, saturated humidity, and a CO2 volume fraction of 5%.

[0066] MTT assay: Tumor cells in the logarithmic growth phase were collected and seeded at 100 μl / well in a 96-well cell culture plate. The plates were incubated at 37°C with 5% CO2 for 24 h to allow cell adhesion. The supernatant was collected, and the required concentration of sample maintenance medium was added. The plates were then incubated at 37°C with 5% CO2 for 48 h. The drug solution was then aspirated, and 20 μL of 5 mg / mL MTT was added. Incubation continued for 4 h. The culture was terminated, and 150 μL of DMSO was added to each well. The plates were shaken for 10 min to fully dissolve and crystallize the cells. The absorbance of each well was measured at 490 nm using a microplate reader. Control wells (cells, drug dissolution medium of the same concentration DMSO, DMSO, MTT) and apoptosis wells (culture medium, DMSO, MTT) were also included.

[0067] The table shows the antitumor activity of the magnolol-magnolol conjugate.

[0068]

[0069] 'a' indicates that the data is the average of three independent experiments (excluding the positive control).

[0070] The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural or procedural transformations made based on the content of the present invention specification, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. A novel magnolol and herb root alkaloid conjugate, characterized in that, Includes the following general structural formula I: Where R is magnolol or a derivative of magnolol, and n is a natural number greater than 1.

2. The novel magnolol and herb root alkaloid conjugate according to claim 1, characterized in that, n is an integer from 2 to 10.

3. A method for preparing a novel magnolol and herb root alkaloid conjugate according to any one of claims 1-2, characterized in that, Includes the following steps: (a) Using acetone or acetonitrile as solvent, magnolol and dibromoalkanes as reactants in a molar ratio of 1:1 to 1.5, react in the presence of potassium carbonate at 60°C or 82°C for 6-12 hours to obtain intermediate B. (b) Using acetonitrile, tetrahydrofuran or N,N-dimethylformamide as solvent, intermediate B and purslane as reactants in a molar ratio of 1:1 to 1.5, the reaction is carried out under reflux at 82°C in the presence of potassium carbonate to obtain the final product of general formula C. (c) Using acetonitrile as solvent, purslane and dibromoalkanes as reactants in a molar ratio of 1:1 to 1.2, the reaction is carried out under reflux at 82°C in the presence of potassium carbonate to prepare intermediate E. (d) Using acetonitrile as solvent, intermediate E and magnolol as reactants in a molar ratio of 1:1, the reaction was carried out under reflux at 82°C in the presence of potassium carbonate to obtain the final product of general formula C.

4. The method for preparing a novel magnolol and herb root alkaloid conjugate according to claim 3, characterized in that, In step (a), the molar ratio of magnolol, dibromoalkanes and potassium carbonate is 1:1.2:1.8, the reaction temperature is 82℃, and the reaction time is 8 hours.

5. The method for preparing a novel magnolol and herb root alkaloid conjugate according to claim 3, characterized in that, In step (b), the molar ratio of intermediate B, purslane and potassium carbonate is 1.2:1:1.5, the solvent is acetonitrile, and the reflux time is 6 hours.

6. The method for preparing a novel magnolol and herb root alkaloid conjugate according to claim 3, characterized in that, In step (c), the molar ratio of purslane, 1,10-dibromodecane and potassium carbonate is 1:1.2:1.5, the solvent is acetonitrile, and the reflux time is 6 hours.

7. The method for preparing a novel magnolol and herb root alkaloid conjugate according to claim 3, characterized in that, In step (d), the molar ratio of intermediate E, magnolol and potassium carbonate is 1:1:1.2, the solvent is acetonitrile, and the reflux time is 6 hours.

8. The application of the novel magnolol and herbaceous root alkaloid conjugate prepared by the method described in any one of claims 1-2 or 3-7, characterized in that, Application of novel magnolol and herb root alkaloid conjugates in the field of antitumor therapy.

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