CN111825615B - A kind of isoaporphine alkaloid derivative and use thereof - Google Patents

Abstract

The invention discloses an oxidized isoaporphine derivative shown as a formula I, wherein R is shown as

A is selected from-O-or a covalent bond, n is an integer of 0-4, R₁Is selected from

‑NR₂R₃；R₂Selected from H, R₃A linear or branched alkyl selected from C1-C3; with the proviso that A is selected from-O-, n ═ 2, 3, R₁Is selected from

Compared with the oxidized isoaporphine, the oxidized isoaporphine alkaloid derivative has obviously improved inhibitory activity and solubility on lung cancer, liver cancer and breast cancer. The invention also discloses the application of the oxidized isoaporphine derivative in preparing antitumor drugs.

Description

A kind of isoaporphine alkaloid derivative and use thereof

technical field

The invention belongs to the fields of medicinal chemistry and medicinal therapy, and particularly relates to an oxidized isoaporphine alkaloid derivative, a preparation method thereof, and the use of the new compound in the preparation of anticancer drugs.

Background technique

Cancer is a major disease affecting human health and longevity, and has now become one of the most important public health problems in the world. According to the "Global Cancer Report", in 2018, there are expected to be 18.1 million new cancer cases and 9.6 million deaths worldwide. The global cancer burden has further increased, and more and more people are paying attention to cancer treatment. Natural products are a treasure trove for the discovery of new anti-tumor drugs. At present, a variety of natural products have been confirmed to have anti-tumor activity, but natural products often have problems such as toxic side effects, solubility, and stability, and they need to be structurally modified to make them become an anticancer drug that can be used clinically.

Oxoisoaporphine refers to a class of isoquinoline alkaloids containing 1-azabenzanthrone skeleton. It was first isolated from Menispermum dauricum DC. In recent years, the anti-tumor activity of oxidized isoapophylline alkaloids has attracted attention. Studies have shown that the backbone 1 of oxidized isoaporphine alkaloids can be inserted into the double helix structure of DNA through π-π stacking, thereby Induces the occurrence of apoptosis mechanism; affects cell proliferation by inhibiting topoisomerase, thereby showing anti-tumor activity.

Oxidized isoaporphine alkaloids and their derivatives are expected to become novel and highly effective antitumor drugs. Appropriate side chain introduction can significantly enhance the activity of such alkaloids.

SUMMARY OF THE INVENTION

The purpose of the present invention is to select the common parent ring of the oxidized isoapophylline alkaloids-1-azabenzanthrone as the lead, and carry out the structural modification of the C-4 position, and obtain a series of active isoapophytine alkaloids. Oxidized isoaporphine alkaloid derivatives with significantly improved phenanthrene and increased solubility.

The purpose of this invention is to realize through the following technical solutions:

A kind of oxidative isoapophiline derivative as shown in formula I:

所示，A选自-O-或共价键，n为0～4的整数，R₁选自

-NR₂R₃；R₂选自H、R₃选自C1-C3的直链或支链烷基；但不包括A选自-O-，n＝2、3，R₁选自

where R is as

As shown, A is selected from -O- or covalent bond, n is an integer from 0 to 4, and R ₁ is selected from

-NR ₂ R ₃ ; R ₂ is selected from H, R ₃ is selected from C1-C3 linear or branched alkyl; but excluding A is selected from -O-, n=2, 3, R ₁ is selected from

但不包括n＝2、3，R₁选自

或所述的A选自共价键，n＝0，R₁选自

Preferably, the A is selected from -O-, n is an integer from 2 to 4, and R ₁ is selected from

But excluding n=2, 3, R ₁ is selected from

Or said A is selected from covalent bonds, n=0, and R ₁ is selected from

More preferably, the A is selected from -O-, n=2, and R ₁ is selected from

Specifically, the compound shown in formula I is selected from the following compounds:

4-[(2-Morpholine)ethoxy]-1-azabenzoxanthrone; 4-[(2-isopropylamino)ethoxy]-1-azabenzoxanthrone; 4-[ 2-(2-Methylpiperidine)ethoxy]-1-azabenzanthracene; 4-[2-(4-hydroxyethylpiperazine)ethoxy]-1-azabenzanthracene Ketone; 4-[(3-Morpholine)propoxy]-1-azabenzanthracene; 4-[3-(2-methylpiperidine)propoxy]-1-azabenzanthracene Ketone; 4-[4-(2-Methylpiperidine)butoxy]-1-azabenzanthracene; 4-[(4-morpholino)butoxy]-1-azabenzanthracene Ketone; 4-[(4-Isopropylamino)butoxy]-1-azabenzoxanthrone; 4-[4-(4-Methylpiperazine)butoxy]-1-azabenzone Anthrone; 4-[4-(4-Hydroxyethylpiperazine)butoxy]-1-azabenzoxanthrone; 4-piperazine-1-azabenzoxanthrone.

Another object of the present invention is to provide the preparation method of the oxidized isoapophene derivative shown in formula I,

但不包括n＝2、3，R₁选自

时，反应路线如下：When A is selected from -O-, n is an integer from 2 to 4, and R ₁ is selected from

But excluding n=2, 3, R ₁ is selected from

, the reaction route is as follows:

include:

Step (1), dissolving 1-azabenzoxanthrone in diethylene glycol, providing a strong alkaline environment with sodium hydroxide, and generating 4-hydroxy-1-azabenzoxanthrone by a hydroxylation reaction with hydrazine hydrate;

所示的二溴烷反应得到中间体；在碳酸钾和催化量的催化剂CTAB存在的条件下，中间体与式R₁H所示的仲胺反应得到氧化异阿朴菲衍生物。Step (2), take 4-hydroxy-1-aza benzoxanthrone as raw material, under the condition that potassium carbonate exists, and formula

The indicated dibromoalkane is reacted to obtain an intermediate; in the presence of potassium carbonate and a catalytic amount of catalyst CTAB, the intermediate is reacted with a secondary amine of formula R ₁ H to obtain an oxidized isoaporphine derivative.

In step (2), the solvent used is selected from the one in DMF, acetone, acetonitrile, toluene, benzene, xylene, 1,4-dioxane, ethyl acetate, dichloromethane, chloroform, tetrahydrofuran or ether. or more.

The molar ratio of the 4-hydroxy-1-azabenzoxanthrone to potassium carbonate and dibromoalkane is 1:1.5:1; the reaction temperature is 0°C to 80°C.

The molar ratio of the intermediate to potassium carbonate and secondary amine is 1:1.5:1.2; the reaction temperature is 0°C to 60°C.

时，反应路线如下：When A is selected from covalent bonds, n=0, and R ₁ is selected from

, the reaction route is as follows:

The preparation method of the oxidized isoapoliphenidate derivative of the invention has mild reaction conditions, low toxicity of used reagents, easily available raw materials, convenient post-processing and high yield.

The oxidative isoaporphine alkaloid derivatives of the present invention are effective for lung cancer cell line A549, liver cancer cell HepG-2, human breast cancer cell line McF-7, MDA-MB-231 and doxorubicin-resistant cell line McF-7/ADR It has excellent antitumor activity, and its activity is significantly higher than that of oxidized isoapophiline. Therefore, another object of the present invention is to provide the use of the oxidized isoaporphine alkaloid derivatives shown in formula I in the preparation of antitumor drugs.

The tumors are lung cancer, liver cancer and breast cancer.

specific implementation

To further illustrate the present invention, a series of examples are set forth below. These examples are illustrative and should not be construed to limit the invention.

Example 1: Synthesis of 4-[(2-morpholine)ethoxy]-1-azabenzanthrone (I ₁ )

The reaction formula is as follows:

4-Hydroxy-1-azabenzoxanthrone (1 eq) was dissolved in 5 mL of anhydrous DMF, K ₂ CO ₃ (1.5 eq) and 1,2-dibromoethane (1 eq) were added, stirred at 80°C overnight, The reaction was stopped, ethyl acetate and saturated brine were added to the reaction solution to extract and remove DMF, and silica gel column chromatography (eluent was petroleum ether: ethyl acetate P/E=3:1) was purified to obtain the intermediate 4-(2-bromo ethoxy)-1-azabenzoxanthrone, then 4-(2-bromoethoxy)-1-azabenzoxanthrone (1eq) was added to the reaction flask, dissolved in 5mL of anhydrous DMF, added K ₂ CO ₃ (1.5eq), CTAB and morpholine (1.2eq), stirred at 60°C overnight, stopped the reaction, extracted to remove DMF, silica gel column chromatography (eluent: petroleum ether: ethyl acetate P/E=5: 1) Purification to obtain compound I ₁ as a yellow solid in a yield of 67%. Compound I ₁ was identified as 4-[(2-morpholine)ethoxy]-1-azabenzanthrone.

ESI-MS: 360.4.[M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.92 (d, J=6Hz, 1H), 8.78 (d, J=6Hz, 1H), 8.64 (d, J=9Hz, 1H), 8.44 ( d,J＝6Hz,1H),8.04(d,J＝6Hz,1H),7.81(t,J＝9Hz,1H),7.69(t,J＝9Hz,1H),7.16(d,J＝9Hz, 1H), 4.42(d, J=6Hz, 2H), 3.78(4H,s), 3.02(t, J=6Hz, 2H), 2.70(4H,s).

Example 2: Synthesis of compound 4-[(2-isopropylamino)ethoxy]-1-azabenzoxanthrone (I ₂ )

The reaction formula is as follows:

Referring to the preparation method of compound _I1 , isopropylamine was used to replace compound morpholine, and other conditions remained unchanged, to obtain target compound _I2 as a yellow solid with a yield of 58%. Compound 12 was identified as 4-[( ₂ -isopropylamino)ethoxy]-1-azabenzanthrone.

ESI-MS: 333.2. [M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.86 (d, J=9Hz, 1H), 8.77 (d, J=6Hz, 1H), 8.48 (d, J=9Hz, 1H), 8.28 ( d,J＝9Hz,1H),7.98(d,J＝6Hz,1H),7.87(t,J＝7.5Hz,1H),7.73(t,J＝7.5Hz,1H),7.60(d,J＝ 9Hz, 1H), 4.6(s, 1H), 3.96(t, J=6Hz, 2H), 3.45(s, 3H), 1.24(d, J=6Hz, 6H).

Example 3: Synthesis of 4-[2-(2-methylpiperidine)ethoxy]-1-azabenzanthrone ( _I3 )

The reaction formula is as follows:

Referring to the preparation method of compound I ₁ , the compound morpholine was replaced with 2-methylpiperidine, and other conditions remained unchanged to obtain the target compound I ₃ as a yellow solid with a yield of 80%. Compound ₁₃ was identified as 4-[2-(2-methylpiperidine)ethoxy]-1-azabenzanthrone.

ESI-MS: 373.2 [M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.96 (d, J=6Hz, 1H), 8.82 (d, J=3Hz, 1H), 8.69 (d, J=6Hz, 1H), 8.48 ( d,J＝3H,1H),8.10(d,J＝3Hz,1H),7.84(t,J＝4.5Hz,1H),7.69(t,J＝4.5Hz,1H),7.23(d,J＝ 3Hz, 1H), 4.43(t, J＝3Hz, 2H), 3.32(d, J＝3Hz, 1H), 3.06(t, J＝3Hz, 2H), 2.53(t, J＝6Hz, J＝9Hz, 2H), 1.72(t, J=6Hz, J=9Hz, 6H), 1.25(d, 3H).

Example 4: Synthesis of 4-[2-( ₄ -hydroxyethylpiperazine)ethoxy]-1-azabenzoxanthrone (I4)

The reaction formula is as follows:

Referring to the preparation method of compound I ₁ , the compound morpholine was replaced with 4-hydroxyethylpiperazine, and other conditions were unchanged, to obtain the target compound I ₄ as a yellow solid with a yield of 85%. Compound ₁₄ was identified as 4-[2-(4-hydroxyethylpiperazine)ethoxy]-1-azabenzanthrone.

ESI-MS: 404.3 [M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.94 (d, J=6Hz, 1H), 8.81 (d, J=6Hz, 1H), 8.67 (d, J=6Hz, 1H), 8.46 ( d,J＝9Hz,1H),8.08(d,J＝6Hz,1H),7.81(t,J＝6Hz,1H),7.67(t,J＝6Hz,1H),7.20(d,J＝9Hz, 1H), 4.44(t, J＝6Hz, 2H), 3.65(t, J＝6Hz, 2H), 3.04(t, J＝6Hz, 2H), 2.74(s, 8H), 2.59(t, J＝6Hz , 2H).

Example ₅ : Synthesis of 4-[(3-morpholino)propoxy]-1-azabenzanthrone (I5)

The reaction formula is as follows:

Referring to the preparation method of compound _I1 , 1,3-dibromopropane was used to replace compound 1,2-dibromoethane, and other conditions remained unchanged to obtain target compound _I5 , yellow solid, yield 55%. Compound ₁₅ was identified as 4-[(3-morpholino)propoxy]-1-azabenzanthrone.

ESI-MS: 375.2. [M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.95 (d, J=9Hz, 1H), 8.80 (d, J=6Hz, 1H), 8.66 (d, J=6Hz, 1H), 8.47 ( d,J＝9Hz,1H),8.08(d,J＝6Hz,1H),7.83(t,J＝7.5Hz,1H),7.69(t,J＝7.5Hz,1H),7.20(d,J＝ 6Hz, 1H), 4.8(t, J＝6Hz, 2H), 3.79(t, J＝4.5Hz, 4H), 2.68(t, J＝7.5Hz, 2H), 2.55(t, J＝4.5Hz, 4H) ),2.21(q,J=7.5Hz,2H).

Example 6: Synthesis of 4-[3-(2-methylpiperidine)propoxy]-1-azabenzoxanthrone (I ₆ )

The reaction formula is as follows:

Referring to the preparation method of compound I ₁ , replace morpholine with 2-methylpiperidine, and replace compound 1,2-dibromoethane with 1,3-dibromopropane, and other conditions remain unchanged, to obtain the target compound _I6 , yellow Solid, 73% yield. Compound ₁₆ was identified as 4-[3-(2-methylpiperidine)propoxy]-1-azabenzanthrone.

ESI-MS: 387.1 [M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.80 (t, J=7.5Hz, 2H), 8.50 (d, J=9Hz, 1H), 8.27 (d, J=9Hz, 1H), 8.03 (d, J=6Hz, 1H), 7.88 (t, J=7.5Hz, 1H), 7.73 (t, J=7.5Hz, 1H), 7.42 (d, J=6Hz, 1H), 4.36 (s, 2H) ),2.69(br,3H),2.10(br,2H),1.62(br,4H),1.32(br,4H),1.10(d,3H).

Example ₇ : Synthesis of 4-[4-(2-methylpiperidine)butoxy]-1-azabenzanthrone (I7)

The reaction formula is as follows:

Referring to the preparation method of compound I ₁ , morpholine was replaced with 2-methylpiperidine, and compound 1,2-dibromoethane was replaced with 1,4-dibromobutane, and other conditions remained unchanged to obtain the target compound I ₇ , Yellow solid, 11% yield. Compound ₁₇ was identified as 4-[4-(2-methylpiperidine)butoxy]-1-azabenzanthrone.

ESI-MS: 401.1 [M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.95 (d, J=6Hz, 1H), 8.79 (d, J=6Hz, 1H), 8.66 (d, J=6Hz, 1H), 8.47 ( d, J=7.5Hz, 1H), 8.10 (d, J=7.5Hz, 1H), 7.83 (t, J=7.5Hz, 1H), 7.69 (t, J=7.5Hz, 1H), 7.18 (d, J＝6Hz, 1H), 4.35(t, J＝6Hz, 2H), 3.86(t, J＝6Hz, 2H), 2.14(t, J＝7.5Hz, 3H), 1.92(t, J＝6Hz, 2H) ),1.32(t,11H).

Example ₈ : Synthesis of 4-[(4-morpholino)butoxy]-1-azabenzanthrone (I8)

The reaction formula is as follows:

Referring to the preparation method of compound _I1 , 1,4-dibromobutane was used to replace compound 1,2-dibromoethane, and other conditions remained unchanged to obtain target compound _I8 , yellow solid, yield 49%. Compound ₁₈ was identified as 4-[(4-morpholino)butoxy]-1-azabenzanthrone.

ESI-MS: 389.2 [M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.93 (d, J=6Hz, 1H), 8.78 (d, J=6Hz, 1H), 8.64 (d, J=6Hz, 1H), 8.44 ( d,J＝6Hz,1H),7.93(t,J＝6Hz,1H),7.82(t,J＝7.5Hz,1H),7.67(d,J＝7.5Hz,1H),7.37(d,J＝ 9Hz, 1H), 4.05(t, J＝6Hz, 2H), 3.33(t, J＝3Hz, 4H), 2.80(t, J＝7.5Hz, 2H), 2.04(t, J＝6Hz, 4H), 1.65(m, 2H), 1.27(m, 2H).

Example ₉ : Synthesis of 4-[(4-isopropylamino)butoxy]-1-azabenzanthrone (I9)

The reaction formula is as follows:

Referring to the preparation method of compound I ₁ , morpholine was replaced with 4-isopropylamino group, and compound 1,2-dibromoethane was replaced with 1,4-dibromobutane, and other conditions remained unchanged to obtain target compound I ₉ , yellow Solid, 19% yield. Compound ₁₉ was identified as 4-[(4-isopropylamino)butoxy]-1-azabenzanthrone.

ESI-MS: 361.3 [M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.94 (d, J=6Hz, 1H), 8.77 (d, J=6Hz, 1H), 8.65 (d, J=9Hz, 1H), 8.45 ( d,J＝6Hz,1H),8.08(d,J＝6Hz,1H),7.81(t,J＝7.5Hz,1H),7.67(t,J＝7.5Hz,1H),7.17(d,J＝ 9Hz, 1H), 4.34(s, 2H), 2.98(m, 1H), 2.48(t, J=6Hz, 2H), 1.91(m, 4H), 1.28(s, 6H).

Example 10: Synthesis of 4-[4-(4-methylpiperazine)butoxy]-1-azabenzoxanthrone (I ₁₀ )

The reaction formula is as follows:

Referring to the preparation method of compound I ₁ , replace morpholine with 4-methylpiperazinyl, and replace compound 1,2-dibromoethane with 1,4-dibromobutane, and other conditions remain unchanged to obtain target compound I ₁₀ , a yellow solid, 12% yield. Compound I ₁₀ was identified as 4-[4-(4-methylpiperazine)butoxy]-1-azabenzanthrone.

ESI-MS: 402.3 [M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.94 (d, J=6Hz, 1H), 8.78 (d, J=9Hz, 1H), 8.64 (d, J=6Hz, 1H), 8.46 ( d,J＝9Hz,1H),8.08(d,J＝6Hz,1H),7.83(t,J＝7.5Hz,1H),7.68(t,J＝7.5Hz,1H),7.16(d,J＝ 6Hz, 1H), 4.34(t, J＝6Hz, 2H), 3.85(t, J＝6Hz, 2H), 2.12(m, J＝6Hz, 2H), 1.93(m, J＝6Hz, 2H), 1.71 (br,3H),1.31(m,8H).

Example 11: Synthesis of 4-[4-(4-hydroxyethylpiperazine)butoxy]-1-azabenzoxanthrone (I ₁₁ )

The reaction formula is as follows:

Referring to the preparation method of compound I ₁ , replace morpholine with 4-hydroxyethylpiperazinyl, replace compound 1,2-dibromoethane with 1,4-dibromobutane, and other conditions remain unchanged, to obtain target compound I ₁₁ , yellow solid, 11% yield. Compound I ₁₁ was identified as 4-[4-(4-hydroxyethylpiperazine)butoxy]-1-azabenzanthrone.

ESI-MS: 432.2 [M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.99 (d, J=6Hz, 1H), 8.81 (d, J=6Hz, 1H), 8.68 (d, J=6Hz, 1H), 8.48 ( d,J＝6Hz,1H),8.13(d,J＝6Hz,1H),7.85(t,J＝7.5Hz,1H),7.70(t,J＝7.5Hz,1H),7.21(d,J＝ 6Hz, 1H), 4.38(t, J＝4.5Hz, 2H), 3.87(t, J＝4.5Hz, 2H), 2.15(t, J＝4.5Hz, 2H), 1.94(t, J＝4.5Hz, 2H), 1.38(m, 12H).

Example 12: Synthesis of 4-piperazine-1-azabenzoxanthrone (Compound I ₁₂ )

The reaction formula is as follows:

Referring to the preparation method of compound I ₁ , using 4-(2-bromoethoxy)-1-azabenzoxanthrone as raw material, replacing compound morpholine with piperazine, and other conditions remaining unchanged, the target compound I ₁₂ was prepared , a yellow solid, 12% yield. Compound I ₁₂ was identified as 4-piperazine-1-azabenzanthrone.

ESI-MS: 316.1 [M+H] ⁺ .

¹ H-NMR (300MHz, CDCl ₃ , ppm), δ _H 8.93 (d, J=9Hz, 1H), 8.77 (d, J=9Hz, 1H), 8.63 (d, J=6Hz, 1H), 8.44 ( d,J＝9Hz,1H),7.93(d,J＝6Hz,1H),7.81(t,J＝7.5Hz,1H),7.66(t,J＝7.5Hz,1H),7.34(d,J＝ 9Hz, 1H), 3.32(t, J=4.5Hz, 4H), 3.23(t, J=3Hz, 4H), 1.27(s, 1H).

Table 1. Substituents of compounds I ₁ -I ₁₂

Example 13: Pharmacological experiments

The antitumor activity test of the oxidized isoapophylline alkaloid derivatives of the present invention was carried out by the tetramethylazole blue colorimetric method (MTT method), and the oxidized isoapophyllite (Oxo) and doxorubicin (Dox) were used as positive controls The target compound and positive control drug were tested against five cancer cell lines (lung cancer cell line A549, liver cancer cell line HepG-2, human breast cancer cell line McF-7, MDA-MB-231 and doxorubicin-resistant cell line McF -7/ADR) inhibitory activity.

Instruments: ultra-clean workbench (SW-CJ-1FD, AIRTECH, Sujing Antai), constant temperature CO ₂ incubator (3111, Thermo, USA), inverted biological microscope (IX71, OLYMPUS, Japan), enzyme-linked immunosorbent assay ( Model 680, BIO-RAD, USA), plate shaker (Kylin-bell lab Instruments), autoclave (YXO.SG41.280, Shanghai Huaxian), centrifuge (SIGMA).

Reagents: DMEM (Keygen), RPMI-1640 (Keygen), Fetal Bovine Serum (GIBCO), Trypsin (SIGMA), DMSO (SIGMA).

Cell lines: human non-small cell lung cancer cell line A549, liver cancer cell HepG-2, human breast cancer cell line McF-7 and MDA-MB-231, adriamycin-resistant cell line McF-7/ADR (all purchased from Jiangsu Kai Base Biotechnology Co., Ltd.). Human non-small cell lung cancer cell line A549, liver cancer cell HepG-2, human breast cancer cell line McF-7 and MDA-MB-231 were cultured in DMEM medium, and adriamycin-resistant cell line McF-7/ADR was cultured in RPMI-1640 base.

Test compounds: Compounds I ₁ -I ₁₂ , isoaporphine oxide, and doxorubicin DMSO, respectively prepared into 10 mM stock solutions, and then diluted with DMEM or RPMI-1640 culture medium to obtain a series of concentrations.

Methods: The cryopreserved cell lines were recovered and cultured in a constant temperature 37°C CO ₂ incubator. The medium was changed once a day, and the cells were plated when they were in a good exponential growth phase. Add 1 mL of 0.25% trypsin digestion solution, digest for 1-2 minutes, observe the cell state under a microscope, when the adherent cells become round and shrink, the digestion solution can be removed, and 1-2 mL of DMEM or RPMI containing 10% fetal bovine serum is added. -1640 medium to prepare cell suspension, count the cells, calculate the amount of cell suspension required according to the number of cells per well 5×10 ⁴ and the total number of wells, inoculate the cell suspension on a 96-well plate, 100 μL Each well was sealed with PBS and placed in a constant temperature 37°C CO ₂ incubator for 24h.

Take the cells in good exponential growth phase, pour the DMEM/RPMI-1640 medium, wash once with PBS, add 1 mL of 0.25% trypsin digestion solution to digest for 1-2 min, and add 1 mL of DMEM medium containing 10% fetal bovine serum or RPMI -1640 culture medium, a cell suspension with a density of 5×10 ⁴ cells/mL was prepared, and inoculated on a 24-well plate (500 μL/well), and cultured in a constant temperature CO ₂ incubator for 24 h. The test compound was added and cultured for 48h, with DMSO as blank control group. MTT reagent was added to a 24-well plate (50 μL/well) and incubated in an incubator for 4 h. The supernatant was discarded, and DMSO (500 μL/well) was added to dissolve. The absorbance value (OD value) was measured at a wavelength of 570 nm using a microplate reader, and the cell inhibition rate and the IC ₅₀ value of the test compound were calculated according to the formula (calculated by GraphPad Prism5 software).

Cell inhibition rate% = (average OD value of blank control group - average OD value of drug group)/average OD value of blank control group × 100%

Table 2. Antiproliferative effects of test compounds on human tumor cell lines

It can be seen from Table 2 that the inhibitory activity of compound I ₁ - compound I ₁₂ on tumor cells is significantly better than that of oxyisoapophene, and the inhibitory activity on different tumor cells has certain selectivity, and the inhibitory activity on human breast cancer cells McF-7 The activity is superior to the inhibitory activity on human lung cancer cell A549, human liver cancer cell Hep-G2, human breast cancer cell MDA-MB-231 and doxorubicin-resistant cell line McF-7/ADR.

The biological activity of the compound decreased with the length of the carbon chain, when n=2 (compound I ₁ -I ₄ ) showed better biological activity, and compound I ₃ (IC ₅₀ 1.32-7.64 μM) inhibited tumor cells The activity is the most prominent. The 4-position substituent on the anthrone ring of the compound has a significant effect on the cytotoxic activity of the compound, and the one substituted with the methylpiperazine ring has the best activity (I ₃ , I ₆ , I ₇ ).

Claims (6)

1. An oxidized isoaporphine derivative represented by formula I:

wherein R is as

A is selected from oxygen or a bond, n is an integer of 0 to 4, R₁Is selected from

-NR₂R₃；R₂Selected from H, R₃A linear or branched alkyl selected from C1-C3; except that A is selected from oxygen, n is 2, 3, and R is₁Is selected from

The case (1).

2. The oxoisoaporphine derivative according to claim 1, wherein A is selected from the group consisting of oxygen, n is an integer of 2 to 4, and R is₁Is selected from

But not including n 2, 3, R₁Is selected from

Or A is selected from a bond, n ═ 0, and R₁Is selected from

The case (1).

3. Oxidised isoaporphine derivatives according to claim 2, characterized in that a is selected from the group consisting of oxygen, n-2, R₁Is selected from

4. Oxidized isoaporphine derivatives according to claim 1, characterized in that they are selected from:

5. use of the oxidized isoaporphine derivative according to any one of claims 1 to 4 for the preparation of an antitumor medicament.

6. The use according to claim 5, wherein said tumor is lung cancer, liver cancer, breast cancer.