A rose alkaloid derivative, its preparation method and application

Abstract

The present invention discloses an ellipticine derivative with a general structural formula (1). The preparation method thereof is as follows: using ellipticine and its derivatives as raw materials, by continuously changing R1, A, a series of compounds are obtained. The synthesized new compounds have a significant inhibitory effect on colorectal tumor cells, and act on the colorectal cancer cell line HT29, wherein IC 50 The minimum is 1.204μM.

Description

Technical Field

[0001] The present invention relates to a medicine and a preparation method and application thereof, and in particular to an ellipticine derivative and a preparation method and application thereof. Background Art

[0002] Colorectal cancer is a common malignant tumor that usually originates from the intestinal mucosa of the colon or rectum. With the changes in lifestyle and genetic factors, the incidence of colorectal cancer has gradually increased worldwide. According to data from the World Health Organization, colorectal cancer is the second most common cancer in the world, accounting for about 15% of all cancer deaths. The occurrence of colorectal cancer is mainly caused by some gene mutations, such as FGFR2, HRAS, BRAF and PIK3CA. In addition, a high-fat, high-sugar and low-fiber diet, lack of exercise and obesity are also closely related to the occurrence of colorectal cancer. At present, the main treatments for colorectal cancer include surgery, drug therapy and radiotherapy, among which single surgical treatment is suitable for patients with carcinoma in situ and early cancer. For patients with advanced cancer, the effect of surgery is unsatisfactory, and postoperative drug and / or radiotherapy adjuvant therapy is required. Drug treatment usually uses antibodies such as bevacizumab, cetuximab and panitumumab, but the disadvantage is that the use of these anti-angiogenic monoclonal antibodies will cause bleeding, arterial and venous thromboembolism to a certain extent; the use of anti-EGFR antibodies such as cetuximab and panitumumab often causes skin reactions and hypomagnesemia.

[0003] Ellipticine is a natural product containing a benzothiophene skeleton. It is mainly extracted from plants of the genus Rosewood and is a colorless to light yellow crystal or powder. Ellipticine has broad application prospects in clinical medicine and drug research. Studies have shown that it has excellent anti-tumor activity. However, it also has obvious defects: ① Inserting into the complementary double helix structure of DNA as a structural analogue, inhibiting the DNA replication and transcription process; ② Inhibiting the activity of DNA topoisomerase II; ③ Inhibiting the oxidation of cytochrome P450 enzymes. Studies have shown that ellipticine derivatives have the use of inhibiting the proliferation, migration and invasion of colorectal cancer both in vivo and in vitro, and screened out the lead derivative I1 with better activity, and the IC of 1.0 against the HT29 cell line 50 The IC value of LO2 for normal cells is around 1.204μM. 50 At 8.763 μM, the IC of Beas 2B 50 At 13.622 μM. Summary of the invention

[0004] Objectives of the invention: The first objective of the present invention is to provide a ellipticine derivative that improves the effect of colorectal tumors; the second objective is to provide a method for preparing the ellipticine derivative; and the third objective is to improve the application of the ellipticine derivative.

[0005] Technical solution: The ellipticine derivative of the present invention has a chemical structure as shown in formula (1):

[0006]

[0007] Wherein, R1 is selected from nitro, aldehyde, benzyl, hydroxyl, thiol, amino, cyano, methoxy, halogen;

[0008] The A group is a five-, six-, or seven-membered saturated or unsaturated heterocyclic ring, or a C3-C5 nitrogen heteroalkyl group;

[0009] In addition, the A group and the aromatic ring may be connected by several alkyl or alkoxy groups, n = 1-3;

[0010] Preferably, the halogen is selected from the group consisting of a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

[0011] Preferably, R1 is nitro, benzyl, hydroxyl, methoxy, or a halogen atom.

[0012] Preferably, the A group is selected from any of the following groups:

[0013]

[0014] Preferably, the ellipticine derivative has any of the following structures:

[0015]

[0016]

[0017] The method for preparing the ellipticine derivative of the present invention comprises the following steps:

[0018] (1) p-Hydroxybenzyl alcohol removes protons under the action of sodium hydroxide and attacks chloroalkane to obtain compound II under the action of phase transfer catalyst.

[0019] (2) Compound II is chlorinated with thionyl chloride at 0°C in tetrahydrofuran as solvent to obtain compound III. (3) 5'-substituted indole is reacted with 2,5-hexanedione in ethanol with p-toluenesulfonic acid as catalyst to obtain compound IV.

[0020] (4) N-Formylaniline is activated with phosphorus oxychloride in o-dichlorobenzene to form an imine intermediate, which further reacts with compound IV to obtain compound V.

[0021] (5) Aminoacetaldehyde diethyl acetal reacts with compound V at high temperature without solvent to obtain compound VI.

[0022] (6) Compound VI is reduced with sodium borohydride in methanol to give compound VII.

[0023] (7) Compound VII is reacted with benzenesulfonyl chloride in dichloromethane to obtain compound VIII.

[0024] (8) Compound VIII is cyclized in dioxane hydrochloride to give compound IX.

[0025] (9) Compound IX reacts with NaH in DMF and further reacts with compound III to obtain compound I.

[0026]

[0027] Wherein, R1 is selected from nitro, aldehyde, benzyl, hydroxyl, thiol, amino, cyano, methoxy, halogen (such as fluorine atom, chlorine atom, bromine atom, iodine atom) and the like.

[0028] The A group is a five-, six- or seven-membered saturated or unsaturated heterocyclic ring, or a C3-C5 nitrogen heteroalkyl group.

[0029] In addition, the A group and the aromatic ring may be connected by several alkyl or alkoxy groups, n = 1-3;

[0030] Preferably, in step (1), the reaction solvent is selected from one or more of acetonitrile, acetone, water, tetrahydrofuran, methanol, N,N-dimethylformamide, and dimethyl sulfoxide; the catalyst acid is selected from inorganic bases such as potassium carbonate, sodium carbonate, sodium bicarbonate, sodium hydroxide, and potassium hydroxide; the brominating agent is tetrabutylammonium bromide; and the reaction temperature is 45 to 80° C. The reaction process is monitored by TLC, and the developing solvent is petroleum ether:ethyl acetate = 1:1.

[0031] Preferably, in step (2), the reaction solvent is selected from one or more of acetonitrile, dichloromethane, chloroform, tetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide or dichlorothionyl; the reaction temperature is 0°C to 60°C; the reaction time is 8 to 15 hours; the reaction progress is monitored by TLC during the reaction, and the developing solvent is dichloromethane:methanol=30:1.

[0032] Preferably, in step (3), the reaction solvent is selected from one or more of methanol, ethanol, n-butanol, acetonitrile, acetic acid, acetone, tetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide or dioxane, and the catalyst acid is selected from one or more of p-toluenesulfonic acid, hydrochloric acid, sulfuric acid, and benzenesulfonic acid. Reflux reaction. The reaction process is monitored by TLC, and the developing solvent is petroleum ether: ethyl acetate = 20:1.

[0033] Preferably, in step (4), the reaction solvent is selected from one or more of benzene, chlorobenzene, toluene, o-dichlorobenzene, and N,N-dimethylformamide; the reaction temperature is 80° C. to 110° C. The reaction progress is monitored by TLC, and the developing solvent is petroleum ether:ethyl acetate = 10:1.

[0034] Preferably, in step (5), the reaction solvent is selected from one or more of benzene, chlorobenzene, toluene, o-dichlorobenzene, N,N-dimethylformamide, dimethyl sulfoxide, or a solvent-free reaction is used; the catalyst is selected from formic acid, acetic acid, p-toluenesulfonic acid hydrochloride, and the reaction temperature is 100° C. to 150° C. The reaction process is monitored by TLC, and the developing solvent is petroleum ether:ethyl acetate = 2:1.

[0035] Preferably, in step (6), the reaction solvent is selected from one or more of methanol, ethanol, dichloromethane, chloroform, tetrahydrofuran or ethyl acetate; the reaction temperature is 0° C. to 35° C. The reaction progress is monitored by TLC, and the developing solvent is petroleum ether:ethyl acetate = 2:1.

[0036] Preferably, in step (7), the reaction solvent is selected from one or more of N,N-dimethylformamide, acetonitrile, acetone, dichloromethane, chloroform, tetrahydrofuran or dimethyl sulfoxide; the base is selected from potassium carbonate, cesium carbonate, sodium carbonate, triethylamine, N,N-dimethylaminopyridine; the reaction temperature is 0°C to 35°C. The reaction process is monitored by TLC, and the developing solvent is petroleum ether:ethyl acetate = 2:1.

[0037] Preferably, in step (8), the reaction solvent is selected from one or more of dioxane, N,N-dimethylformamide, dimethyl sulfoxide or water; the acid is selected from trifluoroacetic acid and hydrochloric acid; the reaction temperature is 80° C. to 110° C. The reaction process is monitored by TLC, and the developing solvent is dichloromethane:methanol=15:1.

[0038] Preferably, in step (9), the reaction solvent is selected from one or more of N,N-dimethylformamide, tetrahydrofuran, dimethyl sulfoxide, dichloromethane, chloroform, N-methylpyrrolidone, and tert-butanol; the base is selected from potassium tert-butoxide, sodium hydride, and tert-butyl lithium; and the reaction temperature is 0°C to room temperature.

[0039] The invention discloses an application of the ellipticine derivative in preparing a drug for treating tumors, wherein the tumor is a colorectal tumor.

[0040] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: (1) The ellipticine derivative, by 1 A series of new compounds were synthesized, which significantly enhanced the inhibitory effect on colorectal tumor cells. The IC 50 The minimum is 1.204 μM; a new ellipticine derivative was designed and synthesized by the preparation method, and the cell viability was detected by the CCK-8 method, and it was found that the compound can significantly inhibit the occurrence and development of colorectal cancer cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1The results of the Clonning Foramation Assay of compound NS2a on HT29 cells are shown in Figure 1, where (A) is the experimental test graph; (B) is the histogram of data calculation.

[0042] Figure 2 Cell Apoptosis Assay test results of compound NS2a on HT29 cells, where (A) is the experimental test graph; (B) is the histogram of data calculation. DETAILED DESCRIPTION

[0043] The technical solution of the present invention is further described below in conjunction with the accompanying drawings.

[0044] Example 1

[0045] The ellipticine derivative of the present invention has a chemical name of 5,11-dimethyl-6-(4-(2-(pyrrolidin-1-yl)ethoxy)benzyl)-6H-pyrido[4,3-b]carbazole, and the synthesis route is as follows:

[0046]

[0047] (1) Synthesis of intermediate Ⅱ1

[0048] Add 40.28 (mmol) of p-hydroxybenzyl alcohol to a sodium hydroxide (88.61 mmol) aqueous solution (45 ml) and stir for 20 minutes. Add toluene (30 ml), compound 1-(2-chloroethyl)pyrrolidine hydrochloride (48.33 mmol) and tetrabutylammonium bromide (2.42 mmol) in sequence. Then heat the solution to reflux (85-90°C), stir vigorously for 2 hours, separate the organic layer, extract the aqueous phase twice with ethyl acetate and combine with the organic layer, wash with water and brine, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure. Purify the residue by silica gel column chromatography to obtain white solid II1 with a yield of 75%. 1 H NMR(300MHz,Chloroform-d)δ7.27–7.15(m,2H),6.92–6.72(m,2H),4.55(s,2H),3.99(t,J=6.0Hz ,2H),3.90(s,1H),2.84(t,J=6.0Hz,2H),2.59(ddt,J=6.8,4.4,2.4Hz,4H),1.78(p,J=3.1Hz,4H).

[0049] (2) Synthesis of intermediate Ⅲ1

[0050] The tetrahydrofuran solution of intermediate II (27.11 mmol) was cooled to 0°C, thionyl chloride (30.35 mmol) was slowly added dropwise, and stirred at room temperature overnight. When TLC showed that the reaction was complete, the precipitated product was filtered, and the filter cake was washed with cyclohexane: tetrahydrofuran = 1:1, and dried in an oven at 80°C to obtain a white solid III1. The yield was 67%, 1 H NMR(300MHz,DMSO-d6)δ11.05(s,1H),7.44–7.36(m,2H),7.06–6.97(m,2H),4.74(s,2H) ,4.37(t,J=5.1Hz,2H),3.56(q,J=5.0Hz,4H),3.10(d,J=9.0Hz,2H),2.04–1.82(m,4H).

[0051] (3) Synthesis of Intermediate IV

[0052] Equimolar amounts of 5-substituted indole 8-12 (10.61 g, 90.57 mmol), 2,5-hexanedione (10.34 g, 90.57 mmol) and p-toluenesulfonic acid (3.45 g, 18.11 mmol) were added to 80 ml of ethanol and refluxed (93 ° C) for 4 hours. The reaction process was monitored by thin layer chromatography. The reaction mixture was evaporated in a vacuum pump, adjusted to alkalinity by adding water, extracted with ethyl acetate, dried by spin drying, and further purified by PE: EA = 20: 1 column chromatography to obtain a white pure product with a yield of 56.66%. 1 H NMR(300MHz,Chloroform-d)δ8.18(d,J=7.9Hz,1H),7.99(s,1H),7.48(d,J=8.1Hz,1H),7.41(t,J=7.6 Hz,1H),7.26(d,J=8.5Hz,1H),7.13(d,J=7.3Hz,1H),6.94(d,J=7.3Hz,1H),2.85(s,3H),2.54(s,3H).

[0053] (4) Synthesis of Intermediate V

[0054] N-formylaniline (9.54ml, 76.97mmol) and POCl3 (6.66ml, 56.45mmol') were dissolved in 150ml o-dichlorobenzene, stirred at room temperature for 40min, and intermediate IV (10.02g, 51.31mmol) compound was added, and heated at 80℃ for 1-3.5h. The reaction was monitored by thin layer chromatography (TLC) in the solvent system (PE:EA=10:1). After the reaction was completed, the reaction mixture was alkalized in sodium acetate (15g, 50ml distilled water) and fully hydrolyzed (excess sodium acetate solution was hydrolyzed for 20-30min until the reaction liquid changed from green to yellow viscous. Filter, wash the filter cake with purified water, extract the filtrate with ethyl acetate three times, dry it with anhydrous sodium sulfate, and combine it with the filter cake by spin drying. Recrystallize with an appropriate amount of toluene (reflux at 105℃ for 10min) to obtain light yellow intermediate V with a yield of 64.96%. 1 H NMR(400MHz,DMSO-d6)δ11.72(s,1H),10.36(s,1H),8.23(d,J=8.0Hz,1H),7.67–7 .58(m,2H),7.48(t,J=7.6Hz,1H),7.26(t,J=7.5Hz,1H),3.10(s,3H),2.56(s,3H).

[0055] (5) Synthesis of intermediate VI

[0056] Add intermediate V (7.01 g, 31.40 mmol) and 13.3 g (4.46 ml, 31.40 mmol) of aminoacetaldehyde diethyl acetal into a round-bottom flask and react for 3 hours in an oil bath at 130°C. Monitor the reaction by TLC (developing agent: petroleum ether: ethyl acetate = 2:1). After the reaction, a yellow gel product is generated, which is intermediate VI. It gradually solidifies during cooling and can be directly used for the next step.

[0057] (6) Synthesis of Intermediate VII

[0058] Add 200 ml of anhydrous ethanol to the intermediate VI, stir under ice bath until dissolved, add sodium borohydride (3.55 g, 93.96 mmol), the reaction solution releases a large number of bubbles and gradually changes from a yellow transparent solution to a beige emulsion. After reacting for 1 hour, add ice to quench, extract the aqueous phase with ethyl acetate, and concentrate to obtain a light yellow solid, which is the intermediate VII. No further purification is required and the next step can be directly carried out.

[0059] (7) Synthesis of Intermediate VIII

[0060] Dissolve the intermediate VII in dichloromethane, add benzenesulfonyl chloride (11.86 ml, 93.93 mmol), and monitor the reaction on a spot plate after 8 to 10 hours (developing solvent is petroleum ether: ethyl acetate = 2:1). After the reaction is completed, dilute the product with dichloromethane, adjust the organic phase to alkaline with saturated sodium carbonate solution, wash with purified water and saturated sodium chloride solution, and dry over anhydrous sodium sulfate overnight. Concentrate, and recrystallize the product with methanol to obtain intermediate VIII with a yield of 67.32%.

[0061] (8) Synthesis of Intermediate IX

[0062] Add 120 ml of dioxane to intermediate IX (10 g, 20.81 mmol), and then add diluted hydrochloric acid (1:1, 10 ml). Reflux the mixture at 106°C for 20 minutes under nitrogen. After cooling to room temperature, add sodium bicarbonate to adjust the system to alkalinity, and 4.47 g of yellow oval ellipticine appears, with a yield of 87.22%. 1 H NMR (400MHz, DMSO-d6) δ11.45(s,1H),9.71(s,1H),8.40(dd,J=19.3,7.0Hz,2H),7.94(d,J=6.0Hz,1H ),7.60–7.56(m,1H),7.56–7.51(m,1H),7.27(ddd,J=8.1,6.9,1.4Hz,1H),3.25(s,3H),2.78(s,3H).

[0063] (9) Synthesis of target product I1

[0064] Stir in anhydrous N,N-dimethylformamide under argon protection to obtain a solution of compound 5,11-dimethylpyrido[4,3-b]carbazole (100 mg, 405.99 μmol), and add sodium hydride (43.84 mg, 1.83 mmol). After the solution is stirred at 0°C for 1 hour, add compound III1 (154.54 mg, 608.98 μmol) and stir at room temperature for 4 hours. When TLC shows that the reaction is complete, add ice water and wash with ethyl acetate. Separate the organic layer, wash with water and brine, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure. Purify the residue by silica gel column chromatography to obtain the target compound I1 as a white solid with a yield of 83%. 1H NMR(300MHz,Chloroform-d)δ9.73(s,1H),8.59–8.47(m,1H),8.40(d,J=7.9Hz ,1H),7.87(d,J=6.2Hz,1H),7.52(t,J=7.8Hz,1H),7.35(t,J=7.6Hz,1H),7.29( s,1H),7.09(d,J=8.2Hz,2H),6.87(d,J=8.2Hz,2H),5.65(s,2H),4.15(t,J=5.8 Hz, 2H), 3.28 (s, 3H), 3.04 (t, J = 5.8Hz, 2H), 2.82 (s, 7H), 1.87 (d, J = 5.5Hz, 4H). 13 C NMR(75MHz,Chloroform-d)δ157.86,149.65,144.89,141.57,140.90,130.37,128.99,127.38,126.8 2,124.02,120.17,116.15,115.05,108.98,108.76,66.16,54.58,54.39,49.11,23.38,14.82,13.42.

[0065] Example 2

[0066] The ellipticine derivative of the present invention has a chemical name of 5,11-dimethyl-6-(4-(2-(piperidin-1-yl)ethoxy)benzyl)-6H-pyrido[4,3-b]carbazole, and its synthesis route is as follows:

[0067]

[0068] (1) Synthesis of intermediate II2

[0069] Add p-hydroxybenzyl alcohol (24.17 mmol) to a sodium hydroxide (53.17 mmol) aqueous solution (45 ml) and stir for 20 minutes. Add toluene (30 ml), compound 1-(2-chloroethyl)piperidinane hydrochloride (29 mmol) and tetrabutylammonium bromide (14.5 mmol) in sequence. Then heat the solution to reflux (85-90°C), stir vigorously for 2 hours, separate the organic layer, extract the aqueous phase twice with ethyl acetate and combine with the organic layer, wash with water and brine, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure. Purify the residue by silica gel column chromatography to obtain white solid II2. Yield 70%, 1H NMR(400MHz,Chloroform-d)δ7.28(d,J=6.6Hz,2H),6.91–6.85(m,2H),4.61(s,2H),4.09(t,J=6.1 Hz, 2H), 2.77 (t, J = 6.1 Hz, 2H), 2.51 (t, J = 5.5 Hz, 4H), 1.62 (p, J = 5.6 Hz, 4H), 1.45 (q, J = 6.2 Hz, 2H).

[0070] (2) Synthesis of intermediate III2

[0071] The tetrahydrofuran solution of intermediate Ⅱ2 (23.16mmol) was cooled to 0℃, thionyl chloride (46.32mmol) was slowly added dropwise, and stirred at room temperature overnight. When TLC showed that the reaction was complete, the precipitated product was filtered, and the filter cake was washed with cyclohexane:tetrahydrofuran = 1:1. After drying in an oven at 80℃, a white solid Ⅲ2 was obtained. The yield was 67%, 1 H NMR(300MHz,Chloroform-d)δ12.49(s,1H),7.33(d,J=8.0Hz,2H),6.88(d,J=8.1Hz,2H),4.57(d,J=8.2Hz,4H),3 .64(d,J=12.0Hz,2H),3.48–3.33(m,2H),2.83(q,J=11.4Hz,2H),2.28(q,J=13.9Hz,2H),1.88(d,J=13.8Hz,2H).

[0072] (3) Synthesis of target product Ⅰ2

[0073] Stir in anhydrous N,N-dimethylformamide under argon protection to obtain a solution of compound 5,11-dimethylpyrido[4,3-b]carbazole (100 mg, 405.99 μmol), and add sodium hydride (38.97 mg, 1.62 mmol). After the solution is stirred at 0°C for 1 hour, add compound III2 (134.56 mg, 487.19 μmol) and stir at room temperature for 4 hours. When TLC shows that the reaction is complete, add ice water and wash with ethyl acetate. Separate the organic layer, wash with water and brine, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure. Purify the residue by silica gel column chromatography to obtain the target compound I2 as a white solid with a yield of 81%. 1H NMR(300MHz,Chloroform-d)δ9.70(s,1H),8.48(d,J=6.2Hz,1H),8.37(d,J=7.8Hz,1H),7. 83(d,J=6.2Hz,1H),7.49(ddd,J=8.3,7.3,1.2Hz,1H),7.37–7.28(m,1H),7.25(d,J=8.1Hz, 1H),7.11–7.00(m,2H),6.88–6.79(m,2H),5.62(s,2H),4.07(t,J=6.0Hz,2H),3.25(s,3H) ,2.78(d,J=6.1Hz,5H),2.52(t,J=5.5Hz,4H),1.61(p,J=5.5Hz,4H),1.43(d,J=3.1Hz,2H). 13 C NMR(75MHz,Chloroform-d)δ157.79,149.67,144.88,140.94,134.53,130.40,128.98,127.38,126.83,124.91,124.0 2,123.74,122.84,120.18,116.13,115.06,108.97,108.74,65.18,57.52,54.81,49.09,25.12,23.59,14.80,13.40.

[0074] Example 3

[0075] The ellipticine derivative of the present invention has a chemical name of 6-(4-(2-(azepine-1-yl)ethoxy)benzyl)-5,11-dimethyl-6H-pyrido[4,3-b]carbazole, and its synthesis route is as follows:

[0076]

[0077] (1) Synthesis of intermediate II3

[0078] Add p-hydroxybenzyl alcohol (24.17 mmol) to a sodium hydroxide (53.17 mmol) aqueous solution (45 ml) and stir for 20 minutes. Add toluene (30 ml), 1-(2-chloroethyl) azacyclohexane hydrochloride (29 mmol) and tetrabutylammonium bromide (14.5 mmol) in sequence. Then heat the solution to reflux (85-90°C), stir vigorously for 2 hours, separate the organic layer, extract the aqueous phase twice with ethyl acetate and combine with the organic layer, wash with water and brine, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure. Purify the residue by silica gel column chromatography to obtain white solid II3. The yield is 67%. 1H NMR(400MHz,Chloroform-d)δ7.27(d,J=8.6Hz,2H),6.91–6.83(m,2H),4.60(s,2H),4.05(t,J=6.2Hz,2H),2.94(t,J=6.2Hz,2H),2.81–

[0079] 2.74(m,4H),2.32(s,1H),1.71–1.56(m,8H).

[0080] (2) Synthesis of intermediate III3

[0081] The tetrahydrofuran solution of intermediate Ⅱ3 (18.16 mmol) was cooled to 0°C, thionyl chloride (36.32 mmol) was slowly added dropwise, and stirred at room temperature overnight. When TLC showed that the reaction was complete, the precipitated product was filtered, and the filter cake was washed with cyclohexane:tetrahydrofuran = 1:1, and dried in an oven at 80°C to obtain a white solid Ⅲ3. Yield 61%, 1 H NMR(300MHz,Chloroform-d)δ12.52(s,1H),7.39–7.29(m,2H),6.96–6.81(m,2H),4.56(s,4H),3.63(q,J=9.5,8.6Hz,2H),3.48(s,2H),3.23–

[0082] 3.04(m,2H),2.19(q,J=9.7Hz,2H),1.95–1.82(m,4H),1.66(dq,J=16.5,8.8,7.2Hz,2H).

[0083] (3) Synthesis of target product Ⅰ3

[0084] Stir in anhydrous N,N-dimethylformamide under argon protection to obtain a solution of compound 5,11-dimethylpyrido[4,3-b]carbazole (100 mg, 405.99 μmol), and add sodium hydride (38.97 mg, 1.62 mmol). After the solution is stirred at 0°C for 1 h, compound III3 (148.23, 487.19 μmol) is added and stirred at room temperature for 4 h. When TLC shows that the reaction is complete, ice water is added and washed with ethyl acetate. The organic layer is separated, washed with water and brine, dried over anhydrous sodium sulfate, filtered, and the solvent is removed under reduced pressure. The residue is purified by silica gel column chromatography to obtain the target compound I3 as a yellow solid with a yield of 83%. 1H NMR(400MHz,Chloroform-d)δ9.69(s,1H),8.47(d,J=6.2Hz,1H),8.37(d,J=7.9Hz,1H),7.83( d,J=6.2Hz,1H),7.54–7.44(m,1H),7.32(t,J=7.5Hz,1H),7.26(d,J=3.5Hz,1H),7.06(d,J=8. 4Hz,2H),6.86–6.79(m,2H),5.63(s,2H),4.23(t,J=5.4Hz,2H),3.25(s,3H),3.19(t,J=5.4Hz ,2H),3.12–2.99(m,4H),2.79(s,3H),1.81(p,J=5.4Hz,4H),1.69–1.60(m,J=5.2,4.4Hz,4H). 13 C NMR(101MHz,Chloroform-d)δ158.07,149.76,144.96,141.60,141.10,134.57,130.24,128.96,127.35,126.81,124.95,1 24.02,123.80,122.91,120.15,116.09,115.10,109.00,108.77,66.04,56.29,55.77,49.17,27.25,27.01,14.83,13.43.

[0085] Example 4

[0086] The ellipticine derivative of the present invention has a chemical name of 2-(4-(5,11-dimethylpyrido[4,3-b]aminooxazol-6-yl)methyl)phenoxy)-N,N-diethyl-1-amine, and its synthesis route is as follows:

[0087]

[0088] (1) Synthesis of intermediate II4

[0089] Add p-hydroxybenzyl alcohol (24.17 mmol) to a sodium hydroxide (53.17 mmol) aqueous solution (45 ml) and stir for 20 minutes. Add toluene (30 ml), 2-chloro-N, N-diethylethane-1-amine (29 mmol) and tetrabutylammonium bromide (14.5 mmol) in sequence. Then heat the solution to reflux (85-90 ° C), stir vigorously for 2 hours, separate the organic layer, extract the aqueous phase twice with ethyl acetate and combine it with the organic layer, wash with water and brine, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure. Purify the residue by silica gel column chromatography to obtain white solid II3. The yield is 55%. 1H NMR(300MHz,Chloroform-d)δ9.88(d,J=2.0Hz,1H),7.88–7.78(m,2H),7.05–6.96(m,2H), 4.13(t,J=6.2Hz,2H), 2.91(t,J=6.3Hz,2H), 2.66(q,J=7.2Hz,4H), 1.08(t,J=7.1Hz,6H).

[0090] (2) Synthesis of intermediate III4

[0091] The tetrahydrofuran solution of intermediate II4 (18.16 mmol) was cooled to 0°C, thionyl chloride (36.32 mmol) was slowly added dropwise, and stirred at room temperature overnight. When TLC showed that the reaction was complete, the precipitated product was filtered, and the filter cake was washed with cyclohexane:tetrahydrofuran = 1:1, and dried in an oven at 80°C to obtain a white solid III3. The yield was 53%. 1 H NMR(300MHz,Chloroform-d)δ7.26(s,2H),6.92–6.83(m,2H),4.61(s,2H),4.05(t ,J=6.2Hz,2H),2.89(t,J=6.4Hz,2H),2.66(q,J=7.2Hz,4H),1.08(t,J=7.4Hz,6H).

[0092] (3) Synthesis of target product Ⅰ4

[0093] Stir in anhydrous N,N-dimethylformamide under argon protection to obtain a solution of compound 5,11-dimethylpyrido[4,3-b]carbazole (100 mg, 405.99 μmol), and add sodium hydride (38.97 mg, 1.62 mmol). After the solution is stirred at 0°C for 1 h, compound III3 (148.23, 487.19 μmol) is added and stirred at room temperature for 4 h. When TLC shows that the reaction is complete, ice water is added and washed with ethyl acetate. The organic layer is separated, washed with water and brine, dried over anhydrous sodium sulfate, filtered, and the solvent is removed under reduced pressure. The residue is purified by silica gel column chromatography to obtain the target compound I4 as a yellow solid with a yield of 87%. 1 H NMR(300MHz,Chloroform-d)δ12.60(s,1H),7.33(d,J=7.9Hz,2H),6.89(d,J=8.1Hz ,2H),4.55(t,J=3.5Hz,4H),3.46(s,2H),3.38–3.12(m,4H),1.47(t,J=7.0Hz,6H).

[0094] Example 5

[0095] The ellipticine derivative of the present invention has a chemical name of 9-bromo-5,11-dimethyl-6-(4-(2-(pyrrolidin-1-yl)ethoxy)benzyl)-6H-pyrido[4,3-b]carbazole, and its synthesis route is as follows:

[0096] The solution of compound 9-bromo-5,11-dimethyl-6H-pyrido[4,3-b]carbazole (100 mg, 307.49 μmol) was obtained by stirring in anhydrous N,N-dimethylformamide under argon protection, and sodium hydride (33.21 mg, 1.38 mmol) was added. After the solution was stirred at 0°C for 1 h, compound III1 (148.23, 487.19 μmol) was added and stirred at room temperature for 4 h. When TLC showed that the reaction was complete, ice water was added and washed with ethyl acetate. The organic layer was separated, washed with water and brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography to obtain the target compound Ⅰ4 as a yellow solid with a yield of 83%. 1 H NMR(400MHz,Chloroform-d)δ9.66(d,J=2.5Hz,1H),8.48(d,J=6.2Hz,1H),8.39(t,J=2.5Hz,1 H),7.81(d,J=6.2Hz,1H),7.55(dd,J=8.7,1.9Hz,1H),7.09(dd,J=8.8,1.7Hz,1H),6.99(d,J=8 .2Hz,2H),6.87–6.80(m,2H),5.54(d,J=3.9Hz,2H),4.06(t,J=5.9Hz,2H),3.14(d,J=4.2Hz,3 H),2.88(t,J=5.9Hz,2H),2.76(d,J=2.4Hz,3H),2.62(q,J=5.9,4.5Hz,4H),1.85–1.74(m,4H). 13 C NMR(101MHz,Chloroform-d)δ158.26,149.85,143.44,141.42,141.35,134.73,129.88,129.57,129.45,126.65,126.4 5,125.42,123.67,122.81,116.05,115.12,112.79,110.28,109.13,66.90,54.98,54.69,49.11,23.45,14.74,13.29.

[0097] Example 6

[0098] The ellipticine derivative of the present invention has a chemical name of 9-methoxy-5,11-dimethyl-6-(4-(2-(pyrrolidin-1-yl)ethoxy)benzyl)-6H-pyrido[4,3-b]carbazole, and its synthesis route is as follows:

[0099] The solution of compound 9-bromo-5,11-dimethyl-6H-pyrido[4,3-b]carbazole (100 mg, 361.87 μmol) was obtained by stirring in anhydrous N,N-dimethylformamide under argon protection, and sodium hydride (39.08 mg, 1.63 mmol) was added. After the solution was stirred at 0°C for 1 h, compound III1 (119.94, 434.25 μmol) was added and stirred at room temperature for 4 h. When TLC showed that the reaction was complete, ice water was added and washed with ethyl acetate. The organic layer was separated, washed with water and brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography to obtain the target compound I6 as a yellow solid with a yield of 87%. 1 H NMR(400MHz,Chloroform-d)δ9.67(s,1H),8.45(d,J=6.2Hz,1H),7.91(d,J=2.3 Hz,1H),7.83(d,J=6.2Hz,1H),7.19–7.11(m,2H),7.06(dd,J=9.0,2.4Hz,2H),6 .89–6.80(m,2H),5.62(s,2H),4.22(t,J=5.4Hz,2H),3.95(s,3H),3.25(s,3H), 3.14(t,J=5.4Hz,2H), 2.97(d,J=6.5Hz,4H), 2.80(s,3H), 1.94(q,J=3.4Hz,4H). 13 C NMR(101MHz,Chloroform-d)δ157.49,154.18,149.58,142.22,140.71,139.72,134.55,130.86,129.19,126.89, 116.14,115.07,114.59,109.39,109.02,108.74,65.49,56.29,54.63,54.59,49.27,23.32,14.75,13.40,1.05.

[0100] Example 7

[0101] The ellipticine derivative of the present invention has a chemical name of 9-benzyloxy-5,11-dimethyl-6-(4-(2-(pyrrolidin-1-yl)ethoxy)benzyl)-6H-pyrido[4,3-b]carbazole, and its synthesis route is as follows:

[0102] Stir in anhydrous N,N-dimethylformamide under argon protection to obtain a solution of compound 9-benzyloxy-5,11-dimethyl-6H-pyrido[4,3-b]carbazole (300 mg, 851.22 μmol), and add sodium hydride (102.14 mg, 4.26 mmol). After the solution was stirred at 0°C for 1 h, compound III1 (282.13, 1.02 mmol) was added and stirred at room temperature for 4 h. When TLC showed that the reaction was complete, ice water was added and washed with ethyl acetate. The organic layer was separated, washed with water and brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography to obtain the target compound Ⅰ7 as a yellow solid with a yield of 34%. 1 H NMR(300MHz,Chloroform-d)δ9.66(s,1H),8.46(d,J=6.2Hz,1H),7.95(d,J=2.3Hz,1H),7 .79(d,J=6.2Hz,1H),7.54–7.49(m,2H),7.44–7.32(m,3H),7.19–7.10(m,2H),7.07–6.99( m,2H),6.89–6.78(m,2H),5.55(s,2H),5.18(s,2H),4.05(t,J=6.0Hz,2H),3.17(s,3H),2. 87 (t, J = 6.0 Hz, 2H), 2.75 (s, 3H), 2.61 (ddt, J = 6.8, 4.2, 2.6 Hz, 4H), 1.79 (p, J = 3.1 Hz, 4H). 13 CNMR(101MHz,Chloroform-d)δ158.19,153.26,149.78,141.10,139.91,137.39,130.26,129.03,128.67,128.04,127.65,126.75, 124.76,124.28,122.63,116.01,115.52,115.05,110.43,109.30,108.68,71.39,67.00,55.03,54.70,49.25,23.48,14.67,13.33.

[0103] Example 8

[0104] The ellipticine derivative of the present invention has a chemical name of 9-nitro-5,11-dimethyl-6-(4-(2-(pyrrolidin-1-yl)ethoxy)benzyl)-6H-pyrido[4,3-b]carbazole, and its synthesis route is as follows:

[0105] The solution of compound 9-nitro-5,11-dimethyl-6H-pyrido[4,3-b]carbazole (194 mg, 665.96 μmol) was obtained by stirring in anhydrous N,N-dimethylformamide under argon protection, and sodium hydride (79.91 mg, 3.33 mmol) was added. After the solution was stirred at 0°C for 1 h, compound III1 (200.73, 799.15 μmol) was added and stirred at room temperature for 4 h. When TLC showed that the reaction was complete, ice water was added and washed with ethyl acetate. The organic layer was separated, washed with water and brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography to obtain the target compound Ⅰ7 as a yellow solid with a yield of 47%. 1 H NMR(400MHz,Chloroform-d)δ9.73(s,1H),9.18(d,J=2.3Hz,1H),8.55(d,J=6.2Hz, 1H),8.38(dd,J=9.0,2.2Hz,1H),7.87(d,J=6.2Hz,1H),7.25(s,1H),7.04(d,J=8.5 Hz,2H),6.88(dd,J=8.8,2.5Hz,2H),5.74(s,2H),4.10(t,J=5.8Hz,2H),3.28(s,3H ), 2.93 (t, J = 5.8Hz, 2H), 2.85 (s, 3H), 2.68 (d, J = 5.9Hz, 4H), 1.82 (p, J = 3.2Hz, 4H). 13 C NMR(101MHz,Chloroform-d)δ158.47,149.98,148.28,142.13,141.73,141.25,135.27,130.20,128.60,12 6.63,123.47,123.34,120.02,116.20,115.35,110.62,108.41,66.84,54.93,54.69,49.45,23.45,13.28.

[0106] application

[0107] 1. CCK-8 assay to detect the inhibitory effect of compound I on colorectal cancer cell lines

[0108] Bazedoxifene was used as a positive control, and the concentration gradient of bazedoxifene and compound I was 0.94, 1.88, 3.75, 7.50, 15.00, and 30.00 μM. The colorectal cancer cell line HT29 was acted on, and the initial cell number was 4000 per well. The cell viability at 48h was directly measured, and the IC of the compound was finally determined. 50 .

[0109] Cell viability = [(experimental group reading - blank group reading) / (control group reading - blank group reading)] × 100%.

[0110] Table 1 Inhibitory effect of compound I on HT29

[0111]

[0112] 2. Inhibitory effect of compound I1 on the growth of HT29 cells

[0113] HT29 cells were seeded into six-well plates at 1000 cells per well and treated with different concentrations of I1 (1 μM, 2 μM, 4 μM, 8 μM and 12 μM) and a 0.1% DMSO control group. After 48 h of treatment, the cells were washed with pre-cooled PBS and cultured for 10-14 days. After colonies were formed, the cells were fixed with 4% paraformaldehyde, stained with 1% crystal violet stain, and counted using Image J. The results are shown in Figure 2. Figure 1 .

[0114] 3. Effect of compound I1 on apoptosis of HT29 cells

[0115] HT29 cells were seeded into 12-well plates at 30w per well, and treated with 2μM and 8μM I1 at different concentrations and a 0.1% DMSO control group. After 48 hours of treatment, the cells were washed with pre-cooled PBS, stained with PI and FITC dyes, and detected by flow cytometry. The results are shown in Figure 2. Figure 2 .

[0116] In summary, based on the effect of bazedoxifene on gp130, we designed and synthesized a series of compounds I. In vitro experiments showed that compound I significantly inhibited the vitality and growth of colorectal cancer cells HT29 and promoted HT29 cell apoptosis, which provided a possibility for the development of small molecule anti-tumor drugs for colorectal cancer and had good application prospects.

Claims

1. An ellipticine derivative having a compound structure as shown in formula (1): in, R1 is selected from nitro, aldehyde, benzyl, hydroxyl, thiol, amino, cyano, methoxy or halogen; Group A is selected from any of the following groups: Furthermore, n=1-3.

2. The ellipticine derivative according to claim 1, characterized in that The R1 is -OH, -NO2, -OCH3, -Br, -OBn or -CN.

3. An ellipticine derivative, characterized in that: The ellipticine derivative has any of the following structures:

4. A method for preparing the ellipticine derivative according to any one of claims 1 to 3, characterized in that: The following steps are involved: (1) p-Hydroxybenzyl alcohol removes protons under the action of sodium hydroxide and attacks chloroalkane to obtain compound II under the action of TBAB; (2) Using tetrahydrofuran as solvent, compound II is chlorinated with thionyl chloride to obtain compound III; (3) The 5'-substituted indole is reacted with 2,5-hexanedione in ethanol using p-toluenesulfonic acid as a catalyst to obtain compound IV; (4) N-formylaniline is activated with phosphorus oxychloride in o-dichlorobenzene to form an imine intermediate, which is further reacted with compound IV to obtain compound V; (5) aminoacetaldehyde diethyl acetal reacts with compound V at high temperature without solvent to obtain compound VI; (6) Compound VI is reduced with sodium borohydride in methanol to obtain compound VII; (7) Compound VII reacts with benzenesulfonyl chloride in dichloromethane to obtain compound VIII; (8) Compound VIII is cyclized in dioxane hydrochloride to obtain compound IX; (9) Compound IX reacts with NaH in DMF and further reacts with compound III to obtain compound I; 5. The method for preparing the ellipticine derivative according to claim 4, characterized in that: In step (2), the reaction solvent is selected from one or more of N,N-dimethylformamide, acetonitrile, acetone, tetrahydrofuran and dimethyl sulfoxide; and the reaction temperature is 0 to 50°C.

6. The method for preparing the ellipticine derivative according to claim 4, characterized in that: In step (3), the reaction solvent is selected from one or more of methanol, ethanol, n-butanol, tert-butanol and propylene glycol.

7. The method for preparing the ellipticine derivative according to claim 4, characterized in that: In step (4), the reaction solvent is selected from one or more of chlorobenzene, o-dichlorobenzene, toluene, o-xylene, and benzene; and the reaction temperature is 80° C. to 100° C.

8. The method for preparing an ellipticine derivative according to claim 4, characterized in that: In step (7), the reaction solvent is selected from one or more of dichloromethane, N,N-dimethylformamide, acetonitrile, acetone, tetrahydrofuran and dimethyl sulfoxide; the base used is potassium carbonate, sodium carbonate, sodium bicarbonate, sodium hydroxide, potassium hydroxide or triethylamine, N,N-dimethylaminopyridine; and the reaction temperature is -10°C to 25°C.

9. The method for preparing the ellipticine derivative according to claim 4, characterized in that: In step (9), the reaction solvent is selected from dichloromethane, N,N-dimethylformamide, tert-butanol, tetrahydrofuran, and N-methylpyrrolidone.

10. Use of the ellipticine derivative according to any one of claims 1 to 3 in the preparation of a drug for treating colorectal cancer.

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