Berberine derivatives, their preparation methods and applications

Abstract

This invention discloses berberine derivatives of formula (I) and formula (II), wherein R1 is H, CH 3， R2 is CH3,CH2CH2Cl, and n is a natural number from 4 to 8. It exhibits good inhibitory effects on cancer cells and can be used as an anticancer drug. This invention also discloses its preparation method.

Description

Technical Field

[0001] This invention relates to berberine derivatives and their application in pharmaceutical manufacturing, belonging to the field of pharmaceutical technology. Background Technology

[0002] Studies have shown that berberine has a certain inhibitory effect on telomerase, can inhibit various cancer cells, and exhibits selective cytotoxicity against both cancer cells and normal cells, while also providing some protection to normal cells. However, berberine has poor lipid and water solubility, low bioavailability, and moderate anticancer efficacy, limiting its clinical application. Triazine compounds contain diazotized amino groups (–N=NN<) in their structure, which can be metabolized by P450 enzymes and converted into alkyl diazonium ions that alkylate guanine-rich telomere DNA and proto-oncogene c-myc DNA. Based on the structural characteristics of berberine and triazine, this invention designs and prepares a berberine derivative to obtain a drug with high anticancer activity. Summary of the Invention

[0003] The purpose of this invention is to provide berberine derivatives that have anticancer effects.

[0004] Another object of the present invention is to provide a method for preparing the above-mentioned berberine derivative.

[0005] Another object of the present invention is to provide the anticancer use of the above-mentioned berberine derivatives.

[0006] The present invention will now be described in detail.

[0007] The berberine derivatives provided by this invention have the general formulas I and II:

[0008]

[0009] In the formula, R1 is H and CH3; R2 is CH3 and CH2CH2Cl; and n is a natural number from 4 to 8.

[0010] The berberine derivatives mentioned above have the following representative structural formulas:

[0011]

[0012] The preparation method of the berberine derivative is as follows:

[0013]

[0014] In the formula, R1 is H and CH3; R2 is CH3 and CH2CH2Cl; and n is a natural number from 4 to 8.

[0015] The application of the berberine derivatives in the preparation of anticancer drugs.

[0016] Beneficial effects: Compared with the prior art, the present invention has the following advantages: By introducing alkyltriazine at the 9-position of berberine, berberine and alkyltriazine can exert a dual anticancer effect, significantly enhancing the anticancer activity of berberine derivatives; by using long-chain fatty diamines to prepare berberine triazine dimers, the lipid solubility of berberine derivatives is effectively improved, thereby increasing the anticancer activity and bioavailability of berberine derivatives.

[0017] The invention is further illustrated by the following embodiments, but it should be noted that the scope of the invention is not limited by these embodiments. Detailed Implementation Example

[0018] Preparation of compound (1)

[0019] 3.7 g (10 mmol) of berberine hydrochloride was placed in an autoclave, dissolved in 20 mL of ethanol, and then 5 mL of concentrated ammonia was added. The mixture was heated for 2-3 h, cooled, evaporated to dryness under reduced pressure, and purified by alumina column chromatography to obtain 9-aminoberberine with a yield of 41%. 1 HNMR (DMSO-d6, 400 MHz) δ: 9.70 (s, 1H), 8.87(s, 1H), 8.16(d, J = 9.1 Hz, 1H), 8.01 (d, J = 9.1 Hz, 1H), 7.75(s, 1H), 7.06(s, 1H), 6.10(s, 2H), 4.90 (t, J =5.5 Hz, 2H), 4.30 (t, J = 5.5 Hz, 2H), 4.04(s, 3H), 4.0(brs, 2H).

[0020] 3.56 g (10 mmol) of 9-aminoberberine was added to a mixture of 10 mL of water and 10 mL of ethanol, stirred, and 20 mL of 15% HCl solution was added. 1.04 g (15 mmol) of NaNO2 solution was added dropwise at 0-5 °C until the solution became clear. Then, 5 mL of ethanol solution of 0.45 g (10 mmol) of dimethylamine was added, and the reaction was continued for 7-8 h. The temperature was raised to room temperature, and 2.9 g of NaAc was added. The solid precipitated, filtered, and purified by silica gel column chromatography to obtain compound (1) with a yield of 43%. 1 H NMR (DMSO-d6, 400 MHz) δ:9.69 (s, 1H), 8.87(s, 1H), 8.14(d, J = 9.1 Hz, 1H), 8.00 (d, J= 9.1 Hz, 1H),7.55(s, 1H), 7.07(s, 1H), 6.11(s, 2H), 4.91 (t, J = 5.6 Hz, 2H), 4.30 (t, J =5.6 Hz, 2H), 4.02(s, 3H), 2.47(s, 6H).

[0021] Example 2

[0022] Preparation of compound (2)

[0023] Compound (2) was prepared by replacing 0.45 g (10 mmol) of dimethylamine with 0.935 g (10 mmol) of chloroethyl methylamine, with other procedures the same as in Example 1, and the yield was 47.3%. 1 H NMR (DMSO-d6, 400 MHz) δ: 9.70 (s, 1H), 8.87(s,1H), 8.15(s, 1H), 8.00 (s, 1H), 7.74(s, 1H), 7.06(s, 1H), 6.11(s, 2H), 4.91(t, J = 5.6 Hz, 2H), 4.31 (t, J = 5.6 Hz, 2H), 4.01(s, 3H), 3.51 (t, J =6.1Hz, 2H), 2.83(t, J = 6.1 Hz, 2H), 2.47(s, 3H).

[0024] Example 3

[0025] Preparation of compound (3)

[0026] Compound (3) was prepared by replacing 0.45 g (10 mmol) of dimethylamine with 0.795 g (10 mmol) of chloroethylamine, with other operations the same as in Example 1, and the yield was 44.9%. 1 H NMR (DMSO-d6, 400 MHz) δ: 9.70 (s, 1H), 8.87(s,1H), 8.16(d, J = 9.1 Hz, 1H), 7.99 (d, J = 9.1 Hz, 1H), 7.54(s, 1H), 7.06(s,1H), 6.11(s, 2H), 4.92 (t, J = 5.6 Hz, 2H), 4.31 (t,J = 5.6 Hz, 2H), 4.02(s,3H), 3.52 (t, J = 6.1Hz, 2H), 2.82(t, J = 6.1 Hz, 2H), 2.03(brs, 1H).

[0027] Example 4

[0028] Preparation of compound (4)

[0029] Add 7.12 g (20 mmol) of 9-aminoberberine to a mixture of 20 mL of water and 20 mL of ethanol, stir, add 40 mL of 15% HCl solution, and add 2.08 g (30 mmol) of NaNO2 solution dropwise at 0-5℃ until the solution becomes clear. Then add 1.16 g (10 mmol). N 1 , N 4 The reaction was continued for 9-10 h with 5 mL of dimethylbutyl-1,4-diamine in ethanol. The mixture was then brought to room temperature, and 5.8 g of NaAc was added. The solid precipitated, filtered, and purified by silica gel column chromatography to obtain compound (1) with a yield of 40.8%. 1 H NMR(DMSO-d6, 400 MHz) δ: 9.70 (s, 2H), 8.88(s, 2H), 8.14(d, J = 9.1 Hz, 2H), 7.99 (d, J = 9.1 Hz, 2H), 7.55 (s, 2H), 7.07(s, 2H), 6.10(s, 4H), 4.91 (t, J = 5.6 Hz, 4H), 4.32 (t, J = 5.6Hz, 4H), 4.03(s, 6H), 2.57(s, 4H), 2.47(s,6H), 1.41(s, 4H).

[0030] Example 5

[0031] Preparation of compound (5)

[0032] Use 1.30 g (10 mmol) N 1 , N 5 -Dimethylbutyl-1,5-diamine replaced 1.16 g (10 mmol) N 1 , N4 -Dimethylbutyl-1,4-diamine, and other procedures were the same as in Example 4, to obtain compound (5) with a yield of 40.3%; 1 H NMR (DMSO-d6,400 MHz) δ: 9.71 (s, 2H), 8.87(s, 2H), 8.16(d, J = 9.1 Hz, 2H), 7.99 (d, J =9.1 Hz, 2H), 7.56(s, 2H), 7.07(s, 2H), 6.11(s, 4H), 4.92 (t, J = 5.6Hz, 4H), 4.30 (t, J = 5.6 Hz, 4H), 4.04(s, 6H), 2.59(s, 4H), 2.47(s, 6H), 1.41(s, 4H), 1.29(s, 2H).

[0033] Example 6

[0034] Preparation of compound (6)

[0035] Use 1.44 g (10 mmol) N 1 , N 6 -Dimethylhexyl-1,6-diamine replaced 1.16 g (10 mmol) N 1 , N 4 -Dimethylbutyl-1,4-diamine, and other procedures were the same as in Example 4, to obtain compound (5) with a yield of 39.2%; 1 H NMR (DMSO-d6,400 MHz) δ: 9.70 (s, 2H), 8.86(s, 2H), 8.15(d, J = 9.1 Hz, 2H), 7.99 (d, J =9.1 Hz, 2H), 7.55(s, 2H), 7.07(s, 2H), 6.12(s, 4H), 4.93 (t, J = 5.6 Hz, 4H), 4.32 (t, J = 5.6 Hz, 4H), 4.05(s, 6H), 2.59(s, 4H), 2.48(s, 6H), 1.41(s, 4H), 1.29(s, 4H).

[0036] Example 7

[0037] Preparation of compound (7)

[0038] Use 1.58 g (10 mmol) N 1 , N 7 -Dimethylheptyl-1,7-diamine replaced 1.16 g (10 mmol) N 1 , N 4 -Dimethylbutyl-1,4-diamine, and other procedures were the same as in Example 4, to obtain compound (7) with a yield of 39.0%; 1 H NMR (DMSO-d6,400 MHz) δ: 9.71 (s, 2H), 8.87(s, 2H), 8.17(s, 2H), 8.01 (s, 2H), 7.55(d, J =8.4 Hz, 2H), 7.07(s, 2H), 6.12(s, 4H), 4.93 (t, J = 5.6 Hz, 4H), 4.29 (t, J =5.6 Hz, 4H), 3.99(s, 6H), 2.59(s, 4H), 2.48(s, 6H), 1.41(s, 4H), 1.29(s, 6H).

[0039] Example 8

[0040] Preparation of compound (8)

[0041] Use 1.72 g (10 mmol) N 1 , N 8 -Dimethyloctyl-1,8-diamine replaced 1.16 g (10 mmol) N 1 , N 4 -Dimethylbutyl-1,4-diamine, and other procedures were the same as in Example 4, to obtain compound (8) with a yield of 38.7%; 1 H NMR (DMSO-d6,400 MHz) δ: 9.71 (s, 2H), 8.87(s, 2H), 8.18(d, J = 9.1 Hz, 2H), 8.00 (d, J =9.1 Hz, 2H), 7.56 (d, J= 8.4 Hz, 2H), 7.07(s, 2H), 6.12(s, 4H), 4.91 (t, J =5.6 Hz, 4H), 4.32 (t, J = 5.6 Hz, 4H), 4.03(s, 6H), 2.59(s, 4H), 2.48(s, 6H), 1.41(s, 4H), 1.29(s, 8H).

[0042] Example 9

[0043] Preparation of compound (9)

[0044] Replace 1.16 g (10 mmol) of 1,8-octanediamine with 1.44 g (10 mmol). N 1 , N 4 -Dimethylbutyl-1,4-diamine, other operations were the same as in Example 4, to obtain compound (9) with a yield of 34.1%; 1 H NMR (DMSO-d6, 400 MHz) δ:9.71 (s, 2H), 8.87(s, 2H), 8.16(d, J = 9.1 Hz, 2H), 8.01 (d, J = 9.1 Hz, 2H), 7.54(d, J = 8.4 Hz, 2H), 7.07(s, 2H), 6.13(s, 4H), 4.93 (t, J = 5.6 Hz, 4H), 4.31 (t, J = 5.6 Hz, 4H), 3.99(s, 6H), 2.59(s, 4H), 2.01(brs, 2H), 1.41(s,4H), 1.29(s, 8H).

[0045] Example 10

[0046] Antitumor activity of compounds (1)-(9)

[0047] Human prostate cancer cells (PC-3), human rectal cancer cells (DU145), human breast cancer cells (MDA-MB-231), human colon cancer cells (HCT-116), and human rectal cancer cells (HT-29) in logarithmic growth phase were selected for the experiment. The concentrations of these cells were adjusted to 5 × 10⁻⁶. 5 / mL, seeded at 100μL per well in a 96-well culture plate. After incubation at 37℃ and 5% CO2 for 24h, 10μmol / L berberine and compounds (1)-(9) were added respectively, and an equal amount of culture medium was added to the blank control group. The culture plate was transferred to a CO2 incubator and cultured for 48h at 37℃, 5% CO2 and saturated humidity. 20μL MTT solution (5g / L) was added to each well of the 96-well culture plate, and the culture plate was transferred to a CO2 incubator and cultured for 4h at 37℃, 5% CO2 and saturated humidity. The culture was then terminated and the culture supernatant in the wells was discarded. For cells growing in suspension, the culture medium in the wells was discarded after centrifugation. 100μL dimethyl sulfoxide was added to each well, shaken for 10min, and after the purple-blue crystals were fully dissolved, the absorbance A value of each group was measured at a wavelength of 490nm using an ELISA reader. The IC50 was calculated using the Mosmann method. 50 Values ​​(Table 1).

[0048]

Claims

1. A berberine derivative, characterized in that, It has general formulas I and II: ， In the formula, R1 is H and CH3; R2 is CH3 and CH2CH2Cl; and n is a natural number from 4 to 8.

2. The berberine derivative according to claim 1, characterized in that, Representative examples are as follows: 。 3. The berberine derivative according to claim 1, characterized in that, The preparation method steps are as follows: ， In the formula, R1 is H and CH3; R2 is CH3 and CH2CH2Cl; and n is a natural number from 4 to 8.

4. The berberine derivative according to claim 1, characterized in that, Applications in the preparation of anticancer drugs.

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