Berberine hydrochloride derivatives and preparation methods and applications thereof

Abstract

The present invention belongs to the field of biomedicine technology, and specifically relates to a class of berberine hydrochloride derivatives and preparation methods and applications thereof. The present invention designs and synthesizes berberine hydrochloride derivatives by changing the benzene ring structure of berberine hydrochloride, and verifies through experiments such as cell proliferation that the compound can effectively inhibit the proliferation of prostate cancer and breast cancer cells, and has significant anti-tumor activity.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a class of berberine hydrochloride derivatives and preparation methods and applications thereof. Background Art

[0002] Berberine, also known as berberine, is an isoquinoline alkaloid derived from the traditional Chinese plant rhubarb. It has long been used as an over-the-counter drug to treat intestinal infections and diarrhea. In recent years, with the continuous deepening of research, the anti-tumor effect of berberine has gradually been recognized, especially as an auxiliary drug for clinical tumor treatment. Berberine has a wide range of anti-tumor effects. Studies have shown that its tumor inhibition spectrum mainly includes lung cancer, breast cancer, nasopharyngeal cancer, gastric cancer, liver cancer and intestinal cancer.

[0003] In recent years, the research on berberine has shifted from in vitro to in vivo. This phenomenon largely indicates that the reproducibility and transformation potential of berberine's anti-tumor effect in in vitro cell models and in vivo animal models have attracted people's attention. Berberine hydrochloride is mostly used in the study of anti-inflammatory activity. It can also inhibit the synthesis of DNA, RNA, protein and fat in tumor cells, and can induce apoptosis of many tumor cells. It has been reported in the study of tumors such as colorectal cancer, prostate cancer, esophageal cancer, and breast cancer. Therefore, it may have broad and important application prospects in cardiovascular system, nervous system diseases, tumors and other diseases, and is increasingly valued.

[0004] In summary, berberine hydrochloride has strong anti-tumor activity. However, berberine hydrochloride is a quaternary ammonium biological salt, and has problems such as poor lipophilicity and low bioavailability. Summary of the invention

[0005] The purpose of the present invention is to change the benzene ring structure of berberine hydrochloride to make it a tertiary amine compound, which is beneficial to improve the bioavailability while further enhancing its anti-tumor activity.

[0006] To achieve the above purpose, the present invention designs a new berberine hydrochloride derivative, and its chemical structure is as follows:

[0007] Wherein, R is o-, m-, or p-bromine or methyl, m-chlorine or amino, and p-ethyl or propyl.

[0008] The preferred chemical names and chemical structural formulas are as follows:

[0009] A: 1-(2-bromophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxo[4,5-g]isoquinolin[3,2-a]isoquinolin; +68.-8-yl)vinyl-1-one

[0010]

[0011] B: 1-(3-bromophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxy[4,5]isoquinolin[3,2-a]isoquinolin-8-yl)ethan-1-one

[0012]

[0013] C: 1-(4-bromophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxo[4,5-g]isoquinolin[3,2-a]isoquinolin-8-yl)vinyl-1-one

[0014]

[0015] D: 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolin[3,2-a]isoquinolin-8-yl)-1-(o-tolyl)ethan-1-one

[0016]

[0017] E: 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolin[3,2-a]isoquinolin-8-yl)-1-(m-tolyl)ethan-1-one

[0018]

[0019] F: 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolin[3,2-a]isoquinolin-8-yl)-1-(p-tolyl)ethan-1-one

[0020]

[0021] G: 1-(3-chlorophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxy[4,5-g]isoquinolino[3,2-a]isoquinolin-8-yl)ethan-1-one

[0022]

[0023] H: 1-(3-aminophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxy[4,5-g]isoquinolino[3,2-a]isoquinolin-8-yl)ethan-1-one

[0024]

[0025] I: 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolin[3,2-a]isoquinolin-8-yl)-1-(4-ethylphenyl)ethane-1-one

[0026]

[0027] J: 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolino[3,2-a]isoquinolin-8-yl)-1-(4-propylphenyl)ethan-1-one

[0028]

[0029] The reaction equation and preparation method steps of berberine hydrochloride are as follows:

[0030]

[0031] Preparation of new berberine hydrochloride derivatives: Weigh a certain amount of base and add it to the alcohol solution, stirring until the base is completely dissolved. Add berberine hydrochloride to a three-necked flask, add the prepared alkaline alcohol solution to the three-necked flask, and stir at 0°C until the berberine hydrochloride is completely dissolved. Slowly add acetophenone with different substituents, stir at 0°C for 30 minutes, and after the reaction solution is clarified, turn it to room temperature for reaction for 2 hours. After the reaction is complete, filter it with suction and wash it repeatedly with methanol solution to obtain a yellow solid. The berberine hydrochloride derivatives are separated by column chromatography.

[0032] Among them, the alcohol is ethanol, methanol, isopropanol; the base is sodium hydroxide or triethylamine.

[0033] Acetophenones with different substituents are: 1-(2-bromophenyl)ethane-1-one, 1-(3-bromophenyl)ethane-1-one, 1-(4-bromophenyl)ethane-1-one, 1-(o-tolyl)ethane-1-one, 1-(m-tolyl)ethane-1-one, 1-(p-tolyl)ethane-1-one, 1-(3-chlorophenyl)ethane-1-one, 1-(3-aminophenyl)ethane-1-one, 1-(4-ethylphenyl)ethane-1-one or 1-(4-propylphenyl)ethane-1-one.

[0034] The molar ratio of berberine hydrochloride: base: acetophenone with different substituents is 1:4 to 5:1.

[0035] The eluent and volume ratio of the eluent used for column chromatography separation in post-treatment were methanol: dichloromethane.

[0036] =1:20.

[0037] The present invention provides a novel berberine hydrochloride derivative which can inhibit the proliferation of prostate cancer and breast cancer cells by inducing cell cycle arrest and apoptosis in prostate cancer and breast cancer cells.

[0038] The advantages of the present invention are:

[0039] (1) Changing the benzene ring structure of berberine hydrochloride has a promoting effect on the activity, which can effectively improve the activity of the new derivatives and have a higher yield and purity.

[0040] (2) Screen out a new berberine hydrochloride derivative that has better biological activity and stronger inhibition of prostate cancer and breast cancer cell proliferation than berberine hydrochloride. DETAILED DESCRIPTION

[0041] The specific implementation modes of the present invention are described in detail below.

[0042] Example 1

[0043] Preparation of 1-(2-bromophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxo[4,5-g]isoquinolin[3,2-a]isoquinolin-8-yl)ethen-1-one

[0044] Weigh sodium hydroxide (0.43g, 10.8mmol) and add it to a 50ml beaker, add 20ml methanol, and stir until the sodium hydroxide is completely dissolved. Weigh berberine hydrochloride (1g, 2.69mmol) and add it to a three-necked flask, add the methanol solution of sodium hydroxide to the three-necked flask, and stir at 0℃ until the berberine hydrochloride is completely dissolved. Slowly add 1-(2-bromophenyl)ethane-1-one (1.07g, 5.38mmol) at 0℃, stir at 0℃ for 30min after the addition, and after the reaction liquid is clarified, turn it to room temperature for reaction for 2h. After the reaction is completed, filter it with suction and wash it repeatedly with methanol solution to obtain a yellow solid. Column chromatography (methanol: dichloromethane = 1:20) gave 1.25 g of 1-(2-bromophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxo[4,5-g]isoquinolin[3,2-a]isoquinolin-8-yl)ethen-1-one in a yield of 87.2%. 1 H NMR (400MHz, CDCl3) δ7.58(d,J=7.6Hz,2H),7.53(s,1H),7.31(t,J=7.5Hz,1H),7.25(m,3H),7.11(s,1H) ,6.85(s,1H),6.73(s,1H),6.00(s,2H),4.25(m,2H),3.92(s,3H),3.85(s,3H),2.80(m,2H),1.76(s,2H). 13C NMR (101MHz, CDCl3) δ183.96,151.02,150.81,147.37,146.12,145.40,145.01,135.92,131.99,129.93,128.51,128.41,127.7 5,126.12,123.32,121.45,120.24,118.83,115.90,106.72,106.39,104.03,100.35,98.93,97.64,59.77,55.55,50.55,28.51.

[0045] Example 2

[0046] Preparation of 1-(3-bromophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxy[4,5]isoquinolino[3,2-a]isoquinolin-8-yl)ethan-1-one

[0047] Weigh sodium hydroxide (0.43g, 10.8mmol) and add it to a 50ml beaker, add 20ml ethanol, and stir until the sodium hydroxide is completely dissolved. Weigh berberine hydrochloride (1g, 2.69mmol) and add it to a three-necked flask, add the ethanol solution of sodium hydroxide to the three-necked flask, and stir at 0℃ until the berberine hydrochloride is completely dissolved. Slowly add 1-(3-bromophenyl)ethane-1-one (1.07g, 5.38mmol) at 0℃, stir at 0℃ for 30min after the addition, and after the reaction liquid is clarified, turn it to room temperature for reaction for 2h. After the reaction is completed, filter it with suction and wash it repeatedly with methanol solution to obtain a yellow solid. Column chromatography (methanol: dichloromethane = 1:20) gave 1.27 g of 1-(3-bromophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxy[4,5-g]isoquinolin[3,2-a]isoquinolin-8-yl)ethylene-1-one with a yield of 88.9%. 1 H NMR (400MHz, DMSO) δ7.85(s,1H),7.76(dd,J=13.6,5.2Hz,2H),7.39(t,J=7.9Hz,1H),7.21(s,1H),6.85(d,J=8.4Hz,1H),6.69(m,2H),5.98( m,3H),5.31(t,J=5.7Hz,1H),3.74(s,3H),3.73(s,3H),3.24(td,J=11.1,5.1Hz,2H),3.18(s,1H),2.91(dd,J=13.9,4.9Hz,1H),2.77(m,2H). 13C NMR(101MHz,DMSO)δ197.02(s),149.46(s),146.89(s),146.29(s),143.13(s),138. 82(s),137.86(s),135.40(s),130.73(d,J=7.8Hz),128.53(s),127.41(s),126.97(s) ),124.93(s),122.41(s),121.92(s),118.29(s),112.49(s),107.77(s),103.94(s), 100.94(s),95.10(s),60.08(s),55.81(d,J=9.4Hz),46.82(s),40.93(s),29.41(s).

[0048] Example 3

[0049] Preparation of 1-(4-bromophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxy[4,5-g]isoquinolin[3,2-a]isoquinolin-8-yl)ethylene-1-one

[0050] Weigh sodium hydroxide (0.43g, 10.8mmol) and add it to a 50ml beaker, add 20ml isopropanol, and stir until the sodium hydroxide is completely dissolved. Weigh berberine hydrochloride (1g, 2.69mmol) and add it to a three-necked flask, add the isopropanol solution of sodium hydroxide to the three-necked flask, and stir at 0℃ until the berberine hydrochloride is completely dissolved. Slowly add 1-(4-bromophenyl)ethane-1-one (1.07g, 5.38mmol) at 0℃, stir at 0℃ for 30min after the addition, and after the reaction liquid is clarified, turn it to room temperature for reaction for 2h. After the reaction is completed, filter it with suction and wash it repeatedly with methanol solution to obtain a yellow solid. Column chromatography (methanol: dichloromethane = 1:20) gave 1.16 g of 1-(4-bromophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxy[4,5-g]isoquinolin[3,2-a]isoquinolin-8-yl)ethen-1-one with a yield of 81.1%. 1 H NMR (300MHz, CDCl3) δ7.70(m,2H),7.46(d,J=8.4Hz,2H),7.11(s,1H),6.76(d,J=8.4Hz,2H),6.53(s,1H),5.93(d,J=1.8Hz,2H),5.9 0(s,1H),5.46(dd,J=7.2,4.2Hz,1H),3.86(s,3H),3.79(s,3H),3.44(m,1H),3.27(m,2H),2.92(dd,J=14.6,4.1Hz,1H),2.66(m,2H).13 C NMR(75MHz, CDCl3)δ197.90(s),150.07(s),147.32(s),146.62(s),143.63(s), 138.51(s),135.92(s),131.59(s),129.93(s),128.88(s),127.83(d,J=12.0Hz ),125.45(s),123.09(s),118.67(s),112.25(s),107.87(s),104.17(s),101.0 5(s),95.15(s),60.78(s),56.10(s),55.75(s),47.79(s),41.01(s),30.32(s).

[0051] Example 4

[0052] Preparation of 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolino[3,2-a]isoquinolin-8-yl)-1-(o-tolyl)ethan-1-one

[0053] Weigh sodium hydroxide (0.43g, 10.8mmol) and add it to a 50ml beaker, add 20ml methanol, and stir until the sodium hydroxide is completely dissolved. Weigh berberine hydrochloride (1g, 2.69mmol) and add it to a three-necked flask, add the methanol solution of sodium hydroxide to the three-necked flask, and stir at 0℃ until the berberine hydrochloride is completely dissolved. Slowly add 1-(o-tolyl)ethane-1-one (0.72g, 5.38mmol) at 0℃, stir at 0℃ for 30min after the addition, and after the reaction liquid is clarified, turn it to room temperature for reaction for 2h. After the reaction is completed, filter it with suction and wash it repeatedly with methanol solution to obtain a yellow solid. Column chromatography (methanol: dichloromethane = 1:30) gave 1.08 g of 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolin[3,2-a]isoquinolin-8-yl)-1-(m-tolyl)ethane-1-one with a yield of 85.5%. 1H NMR (400MHz, DMSO) δ7.60(d,J=7.6Hz,1H),7.36(t,J=7.4Hz,1H),7.24(d,J=6.3Hz,2H),7.16(t,J =7.5Hz,1H),6.84(d,J=8.4Hz,1H),6.71(t,J=4.1Hz,2H),6.01(s,1H),5.99(s,1H),5.98(s,1H),5 .35(dd,J=7.2,3.9Hz,1H),3.75(s,3H),3.74(s,3H),3.44(dd,J=15.1,7.3Hz,1H),3.23(dt,J=9. 0,4.4Hz,1H),3.18(m,1H),2.75(t,J=5.5Hz,1H),2.69(ddd,J=14.5,9.0,4.3Hz,2H),2.39(s,3H). 13 C NMR(101MHz,DMSO)δ200.99(s),149.59(s),146.85(s),146.29(s),143.09(s),137.75( d,J=12.1Hz),137.04(s),131.74(s),131.44(s),129.25(s),128.51(s),127.43(s),125 .66(s),124.94(s),123.14(s),118.25(s),112.27(s),107.78(s),103.91(s),100.93( s),94.96(s),60.06(s),55.84(s),55.01(s),47.06(s),43.60(s),29.48(s),21.03(s).

[0054] Example 5

[0055] Preparation of 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolino[3,2-a]isoquinolin-8-yl)-1-(m-tolyl)ethan-1-one

[0056] Weigh sodium hydroxide (0.43g, 10.8mmol) and add it to a 50ml beaker, add 20ml methanol, and stir until the sodium hydroxide is completely dissolved. Weigh berberine hydrochloride (1g, 2.69mmol) and add it to a three-necked flask, add the methanol solution of sodium hydroxide to the three-necked flask, and stir at 0℃ until the berberine hydrochloride is completely dissolved. Slowly add 1-(m-tolyl)ethane-1-one (0.72g, 5.38mmol) at 0℃, stir at 0℃ for 30min after the addition, and after the reaction liquid is clarified, turn it to room temperature for reaction for 2h. After the reaction is completed, filter it with suction and wash it repeatedly with methanol solution to obtain a yellow solid. Column chromatography (methanol: dichloromethane = 1:30) gave 1.11 g of 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolin[3,2-a]isoquinolin-8-yl)-1-(m-tolyl)ethane-1-one with a yield of 87.9%. 1 H NMR (400MHz, DMSO) δ7.56 (m, 2H), 7.33 (dt, J = 15.1, 7.5Hz, 2H), 7.25 (s, 1H), 6.86(d,J=8.4Hz,1H),6.68(m,2H),6.03(s,1H),5.99(d,J=4.4Hz,2H),5.36 (dd,J=7.2,4.2Hz,1H),3.74(s,3H),3.72(s,3H),3.47(dd,J=14.3,7.4Hz,1 H),3.23(dt,J=9.0,4.4Hz,1H),3.18–3.07(m,1H),2.78(m,3H),2.28(s,3H). 13 C NMR(101MHz,DMSO)δ198.16(s),149.51(s),146.85(s),146.28(s),143.10(s),137. 82(d,J=15.7Hz),136.82(s),133.56(s),128.89–128.31(m),127.49(s),125.10(d, J=17.8Hz),122.91(s),118.24(s),112.36(s),107.77(s),103.96(s),100.93(s),9 4.98(s),60.08(s),55.85(s),55.48(s),47.08(s),40.93(s),29.43(s),20.80(s).

[0057] Example 6

[0058] Preparation of 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolino[3,2-a]isoquinolin-8-yl)-1-(p-tolyl)ethan-1-one

[0059] Weigh 1.5 ml of triethylamine and add it to a 50 ml beaker, add 20 ml of methanol, and stir to mix the triethylamine and methanol evenly. Weigh (1 g, 2.69 mmol) of berberine hydrochloride and add it to a three-necked flask, add the mixed solution of triethylamine and methanol to the three-necked flask, and stir at 0°C until the berberine hydrochloride is completely dissolved. Slowly add 1-(p-tolyl)ethane-1-one (0.72 g, 5.38 mmol) at 0°C, stir at 0°C for 30 minutes after the addition, and after the reaction solution is clarified, turn it to room temperature for reaction for 2 hours. After the reaction is completed, filter it with suction and wash it repeatedly with methanol solution to obtain a yellow solid. Column chromatography (methanol: dichloromethane = 1:30) gave 1.16 g of 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolin[3,2-a]isoquinolin-8-yl)-1-(p-tolyl)ethane-1-one in a yield of 91.8%. 1 H NMR (400MHz, DMSO) δ7.74(d,J=8.2Hz,2H),7.24(d,J=7.1Hz,3H),6.86(d,J=8.4Hz ,1H),6.74(d,J=8.4Hz,1H),6.70(s,1H),6.04(s,1H),5.99(d,J=3.5Hz,2H),5.37 (dd,J=7.3,4.3Hz,1H),3.75(s,3H),3.72(s,3H),3.48(dd,J=14.3,7.5Hz,1H),3. 22(dt,J=9.1,4.5Hz,1H),3.13(td,J=10.7,4.0Hz,1H),2.77(m,3H),2.33(s,3H). 13 C NMR(101MHz,DMSO)δ197.57(s),149.50(s),146.84(s),146.28(s),143.35(s),14 3.08(s),137.69(s),134.27(s),129.15(s),128.51(s),128.20(s),127.47(s),12 4.99(s),122.95(s),118.24(s),112.36(s),107.77(s),103.95(s),100.93(s),94 .94(s),60.05(s),55.86(s),55.39(s),47.09(s),40.75(s),29.44(s),21.08(s).

[0060] Example 7

[0061] Preparation of 1-(3-chlorophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxy[4,5-g]isoquinolino[3,2-a]isoquinolin-8-yl)ethan-1-one

[0062] Weigh sodium hydroxide (0.43g, 10.8mmol) and add it to a 50ml beaker, add 20ml methanol, and stir until the sodium hydroxide is completely dissolved. Weigh berberine hydrochloride (1g, 2.69mmol) and add it to a three-necked flask, add the methanol solution of sodium hydroxide to the three-necked flask, and stir at 0℃ until the berberine hydrochloride is completely dissolved. Slowly add 1-(3-chlorophenyl)ethane-1-one (0.83g, 5.38mmol) at 0℃, stir at 0℃ for 30min after the addition, and after the reaction liquid is clarified, turn it to room temperature for reaction for 2h. After the reaction is completed, filter it with suction and wash it repeatedly with methanol solution to obtain a yellow solid. Column chromatography (methanol: dichloromethane = 1:20) gave 1.08 g of 1-(3-chlorophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxy[4,5-g]isoquinolin[3,2-a]isoquinolin-8-yl)ethan-1-one with a yield of 81.9%. 1 H NMR (400MHz, DMSO) δ7.73(dd,J=7.3,4.9Hz,2H),7.62(d,J=8.0Hz,1H),7.46(t,J=7.8Hz,1H),7.21(s,1H),6.85(d,J=8.4Hz,1H),6.68(m,2H), 6.01(s,1H),5.99(s,2H),5.32(t,J=5.7Hz,1H),3.74(s,3H),3.73(s,3 H),3.29(m,2H),3.18(s,1H),2.91(dd,J=13.9,4.8Hz,1H),2.78(m,2H). 13C NMR(101MHz,DMSO)δ197.08(s),149.47(s),146.89(s),146.29(s),143.15(s),138.6 2(s),137.85(s),133.44(s),132.52(s),130.45(s),128.53(s),127.87(s),127.42(s) ),126.62(s),124.94(s),122.42(s),118.29(s),112.49(s),107.77(s),103.95(s), 100.94(s),95.10(s),60.06(s),55.80(d,J=11.6Hz),46.83(s),40.94(s),29.41(s).

[0063] Example 8

[0064] Preparation of 1-(3-aminophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxy[4,5-g]isoquinolino[3,2-a]isoquinolin-8-yl)ethan-1-one

[0065] Weigh sodium hydroxide (0.43g, 10.8mmol) and add it to a 50ml beaker, add 20ml methanol, and stir until the sodium hydroxide is completely dissolved. Weigh berberine hydrochloride (1g, 2.69mmol) and add it to a three-necked flask, add the methanol solution of sodium hydroxide to the three-necked flask, and stir at 0℃ until the berberine hydrochloride is completely dissolved. Slowly add 1-(3-aminophenyl)ethane-1-one (0.73g, 5.38mmol) at 0℃, stir at 0℃ for 30min after the addition, and after the reaction liquid is clarified, turn it to room temperature for reaction for 2h. After the reaction is completed, filter it with suction and wash it repeatedly with methanol solution to obtain a yellow solid. Column chromatography (methanol: dichloromethane = 1:13) gave 1.14 g of 1-(3-aminophenyl)-2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxy[4,5-g]isoquinolin[3,2-a]isoquinolin-8-yl)ethan-1-one in a yield of 90.1%. 1H NMR (400MHz, DMSO) δ7.28(s,1H),7.07(d,J=8.5Hz,2H),7.02(d,J=7.2Hz,1H),6.87( d,J=8.3Hz,1H),6.75(d,J=7.8Hz,2H),6.70(s,1H),6.06(s,1H),5.99(s,2H),5.40( d,J=3.4Hz,1H),5.30(s,2H),3.76(s,3H),3.73(s,3H),3.53(dd,J=14.6,7.6Hz,1H) ,3.21(t,J=11.6Hz,1H),3.10(t,J=8.5Hz,1H),2.68(t,J=16.0Hz,2H),2.60(m,1H). 13 C NMR(101MHz,DMSO)δ198.23(s),149.55(s),149.00(s),146.86(s),146.32(s),143.0 3(s),137.53(d,J=15.8Hz),129.04(s),128.55(s),127.48(s),124.98(s),123.26(s) ),118.48(s),118.22(s),115.81(s),112.65(s),112.27(s),107.81(s),103.95(s), 100.93(s),94.87(s),60.07(s),55.85(s),55.01(s),47.26(s),40.93(s),29.50(s).

[0066] Example 9

[0067] Preparation of 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolino[3,2-a]isoquinolin-8-yl)-1-(4-ethylphenyl)ethan-1-one

[0068] Weigh sodium hydroxide (0.43g, 10.8mmol) and add it to a 50ml beaker, add 20ml methanol, and stir until the sodium hydroxide is completely dissolved. Weigh berberine hydrochloride (1g, 2.69mmol) and add it to a three-necked flask, add the methanol solution of sodium hydroxide to the three-necked flask, and stir at 0℃ until the berberine hydrochloride is completely dissolved. Slowly add 1-(4-ethylphenyl)ethane-1-one (0.80g, 5.38mmol) at 0℃, stir at 0℃ for 30min after the addition, and after the reaction liquid is clarified, turn it to room temperature for reaction for 2h. After the reaction is completed, filter it with suction and wash it repeatedly with methanol solution to obtain a yellow solid. Column chromatography (methanol: dichloromethane = 1:25) gave 1.12 g of 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolin[3,2-a]isoquinolin-8-yl)-1-(4-ethylphenyl)ethane-1-one, with a yield of 86.2%. 1 H NMR (400MHz, DMSO) δ7.66 (m, 2H), 7.33 (dt, J = 15.1, 7.5Hz, 2H), 7.25 (s, 1H) ,6.86(d,J=8.4Hz,1H),6.75(m,2H),6.03(s,1H),5.99(d,J=4.4Hz,2H),5. 36(dd,J=7.2,4.2Hz,1H),3.74(s,3H),3.72(s,3H),3.47(dd,J=14.3,7.4H z,1H),3.23(dt,J=9.0,4.4Hz,1H),3.18(m,1H),2.78(m,3H),2.28(s,3H). 13 C NMR(101MHz,DMSO)δ198.16(s),149.51(s),146.85(s),146.28(s),143.10(s),13 7.82(d,J=15.7Hz),136.82(s),133.56(s),128.89(m),127.49(s),125.10(d,J=1 7.8Hz),122.91(s),118.24(s),112.36(s),107.77(s),103.96(s),100.93(s),94 .98(s),60.08(s),55.85(s),55.48(s),47.08(s),40.93(s),29.43(s),20.80(s).

[0069] Example 10

[0070] Preparation of 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolino[3,2-a]isoquinolin-8-yl)-1-(4-propylphenyl)ethan-1-one

[0071] Weigh sodium hydroxide (0.43g, 10.8mmol) and add it to a 50ml beaker, add 20ml ethanol, and stir until the sodium hydroxide is completely dissolved. Weigh berberine hydrochloride (1g, 2.69mmol) and add it to a three-necked flask, add the ethanol solution of sodium hydroxide to the three-necked flask, and stir at 0℃ until the berberine hydrochloride is completely dissolved. Slowly add 1-(4-propylphenyl)ethane-1-one (0.87g, 5.38mmol) at 0℃, stir at 0℃ for 30min after the addition, and after the reaction liquid is clarified, turn it to room temperature for reaction for 2h. After the reaction is completed, filter it with suction and wash it repeatedly with methanol solution to obtain a yellow solid. Column chromatography (methanol: dichloromethane = 1:30) gave 1.17 g of 2-(9,10-dimethoxy-5,8-dihydro-6H-[1,3]dioxa[4,5]isoquinolin[3,2-a]isoquinolin-8-yl)-1-(4-propylphenyl)ethane-1-one with a yield of 87.4%. 1 H NMR (400MHz, DMSO) δ7.88(d,J=8.2Hz,1H),7.75(d,J=8.2Hz,2H),7.33(t,J=7.8H z,1H),7.24(d,J=8.6Hz,3H),6.86(d,J=8.4Hz,1H),6.68(m,2H),6.04(s,1H),5.9 9(s,2H),5.36(dd,J=7.2,4.4Hz,1H),4.13(s,1H),3.74(s,3H),3.71(s,3H),3.4 9(dd,J=14.3,7.4Hz,1H),2.77(m,4H),1.57(dd,J=14.2,6.7Hz,2H),0.85(m,3H). 13C NMR(101MHz,DMSO)δ197.58(s),149.49(s),147.78(s),146.83(s),146.27(s),143.09(s),1 37.72(s),134.53(s),128.56(d,J=8.4Hz),128.21(s),127.48(s),125.00(s),122.91(s),1 18.22(s),112.36(s),107.75(s),103.94(s),100.92(s),94.95(s),60.03(s),55.85(s),55 .39(s),48.58(s),47.08(s),40.65(s),37.05(s),29.43(s),26.60(s),23.66(s),13.51(s).

[0072] Embodiment 11

[0073] Tumor cell inhibitory activity

[0074] 1. Experimental drugs

[0075] Different concentrations of berberine hydrochloride derivatives AJ (0.5, 1, 2, 4, 8, 16, 32 μM)

[0076] 2. Cell lines

[0077] Human prostate cancer cells (22Rv1) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. They were routinely cultured in 1640 medium containing 10% fetal bovine serum at 37°C, 5% CO2, in an incubator. The medium was replaced every 48 hours. When the cells reached saturation, they were digested and passaged with 0.25% trypsin. The cells were passaged once every 2-3 days. Cells in the logarithmic growth phase were selected for the experiment.

[0078] 3. Cell Viability Detection

[0079] The 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT) method was used to detect the effects of different concentrations of berberine hydrochloride derivatives (AJ) on the proliferation activity of human prostate cancer cell 22RV1. Prostate cancer cells 22Rv1 in the logarithmic phase were taken, digested with trypsin and resuspended, and the cell concentration was adjusted to 5×104 / ml. 100μL / well was inoculated into 96-well plates. After 12h of adherent growth, the culture medium was aspirated and different concentrations of berberine hydrochloride derivatives (AJ) were added, and the corresponding blank control group was set. Three replicate wells were set for each concentration. The cells were cultured in a 37°C, 5% CO2 incubator for 24 h, the supernatant was discarded, and then 20 μL of 5 mg / ml MTT was added and incubated at 37°C, 5% CO2 for 4 h. The supernatant was discarded, 150 μL of DMSO was added to each well, and the cells were shaken for 10 min. The OD value was detected at 490 nm by an enzyme reader, and the half inhibitory concentration (IC 50 ).

[0080] The results showed that berberine hydrochloride derivatives A and C had a strong inhibitory effect on the proliferation of 22Rv1 cells in a concentration-dependent manner. 50 is 0.887, while the IC of derivatives A and C 50 The values ​​of derivatives B and F were 0.814 and 0.4249, respectively, which showed better inhibitory effect on 22Rv1 cell proliferation. 50 The inhibitory effects of berberine hydrochloride and berberine hydrochloride derivatives on 22RV1 cell proliferation are shown in Table 1.

[0081] Table 1. Inhibitory activity of berberine hydrochloride and berberine hydrochloride derivatives on 22RV1 cell proliferation

[0082]

[0083] Berberine hydrochloride derivatives A and C have a strong inhibitory effect on the proliferation of 4T1 cells, and are concentration-dependent. 50 is 1.131, while the IC of derivatives A and C 50 The values ​​were 1.083 and 0.6139, respectively, showing better inhibitory effect on 4T1 cell proliferation. The IC values ​​of derivatives B, F, G, H, and J were 50 The inhibitory effects of berberine hydrochloride and berberine hydrochloride derivatives on 4T1 cell proliferation are shown in Table 2.

[0084] Table 2. Inhibitory activity of berberine hydrochloride and berberine hydrochloride derivatives on 4T1 cell proliferation

[0085]

[0086]

Claims

1. An application of a berberine hydrochloride derivative, characterized in that: The berberine hydrochloride derivative is used to prepare a drug for inhibiting the proliferation of prostate cancer cells or breast cancer cells. The chemical structural formula of the berberine hydrochloride derivative is as follows: 。

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