A trisubstituted acridine derivative and its preparation method and application

Abstract

The present invention belongs to the technical field of acridine derivatives, and provides a trisubstituted acridine derivative and a preparation method and application thereof. The structural formula of the trisubstituted acridine derivative of the present invention is:. The trisubstituted acridine derivative of the present invention has excellent selectivity and stability for quadruple helix structure (G4-DNA), and has a broad spectrum of anti-tumor activity, including but not limited to ovarian cancer, prostate cancer, colorectal cancer, lung cancer, liver cancer, gastric cancer, etc., and has low toxic side effects on normal cells and tissues.

Description

Technical Field

[0001] The present invention relates to the technical field of acridine derivatives, and in particular to a trisubstituted acridine derivative and a preparation method and application thereof. Background Art

[0002] Currently, cancer is a disease with a high mortality rate, and the incidence rate is continuing to rise, and the patients are becoming younger. Drug therapy is currently the main treatment strategy for patients with mid- and late-stage cancer. Although it has achieved certain success in the treatment of certain cancers, it still has huge limitations, such as the serious toxic side effects of drugs on normal cells, the drug resistance of cancer cells to drugs, and the unique heterogeneity of cancer cells. Therefore, finding effective anti-cancer drugs has become an urgent task for public health and medical research.

[0003] Acridine is a nitrogen-containing heterocyclic compound with a tricyclic aromatic structure. Acridine derivatives obtained by modifying different chemical groups can exhibit various biological activities. Acridine derivatives have become potential anti-tumor drug ingredients due to their strong affinity for nucleic acids, especially their interaction with DNA and RNA. At present, acridine derivatives as anti-tumor drugs still have certain defects and challenges, mainly manifested in their lack of selectivity for tumor cells and normal cells, and serious toxic side effects. The quadruple helix structure (G4-DNA) can be clearly distinguished from the conventional double helix DNA structure, and G4-DNA is widely present in the promoter region of proto-oncogenes, making it a potential new target for anti-tumor drugs. Acridine derivatives obtained by modifying different chemical groups are expected to show high activity only to G4-DNA, effectively improving the selectivity of acridine derivatives for tumor cells and reducing their toxic side effects.

[0004] Therefore, it is of great significance to study trisubstituted acridine derivatives with high antitumor activity and selectivity and low toxic side effects targeting G4-DNA. Summary of the invention

[0005] The purpose of the present invention is to provide a trisubstituted acridine derivative and a preparation method and application thereof in view of the deficiencies in the prior art.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides a tri-substituted acridine derivative, the structural formula of the tri-substituted acridine derivative is:

[0008] .

[0009] The present invention also provides a method for preparing the trisubstituted acridine derivative, comprising the following steps:

[0010] 1) condensing 2-chloro-4-aminobenzoic acid and acetic anhydride to obtain a condensation intermediate;

[0011] 2) Mix the condensation intermediate, m-aminoacetanilide, copper powder, cuprous oxide, potassium carbonate and ethylene glycol ether, and perform Ullmann reaction to obtain a benzoic acid intermediate;

[0012] 3) The benzoic acid intermediate and sulfuric acid are subjected to a cyclization reaction to obtain the acridone intermediate 1;

[0013] 4) subjecting the acridone intermediate 1 to a condensation reaction with butyric anhydride to obtain the acridone intermediate 2;

[0014] 5) The acridone intermediate 2 is subjected to an electrophilic substitution reaction with phosphorus oxychloride to obtain an acridine intermediate;

[0015] 6) Mixing the acridine intermediate, N,N-dimethyl-p-phenylenediamine and phenol, and performing a nucleophilic substitution reaction to obtain a trisubstituted acridine intermediate 1;

[0016] 7) hydrolyzing the trisubstituted acridine intermediate 1 with sulfuric acid to obtain the trisubstituted acridine intermediate 2;

[0017] 8) The trisubstituted acridine intermediate 2,3-chloropropionyl chloride, triethylamine and N,N-dimethylformamide are mixed and subjected to a nucleophilic addition reaction to obtain a trisubstituted acridine derivative.

[0018] Preferably, in step 1), the molar volume ratio of 2-chloro-4-aminobenzoic acid to acetic anhydride is 110-120 mmol:30 mL; the temperature of the condensation reaction is 90-110° C., and the time of the condensation reaction is 20-40 min.

[0019] Preferably, in step 2), the molar ratio of the condensation intermediate, m-aminoacetanilide, copper powder, cuprous oxide and potassium carbonate is 1:0.8~1.2:0.08~0.12:0.08~0.12:0.2~0.8, and the molar volume ratio of the condensation intermediate and ethylene glycol ethyl ether is 1mmol:1~2.5mL; the temperature of the Ullmann reaction is 125~145°C, and the time of the Ullmann reaction is 4~6h.

[0020] Preferably, in step 3), the molar volume ratio of the benzoic acid intermediate and sulfuric acid is 14-17 mmol:12 mL, and the mass concentration of sulfuric acid is 92-98%; the temperature of the cyclization reaction is 125-145° C., and the time of the cyclization reaction is 2-4 h.

[0021] Preferably, in step 4), the molar volume ratio of the acridone intermediate 1 to butyric anhydride is 0.8-2.5 mmol:10 mL; the temperature of the condensation reaction is 90-110° C., and the time of the condensation reaction is 1-3 h.

[0022] Preferably, in step 5), the molar volume ratio of the acridone intermediate 2 to phosphorus oxychloride is 12-15 mmol:20 mL; the temperature of the electrophilic substitution reaction is 80-100° C., and the time of the electrophilic substitution reaction is 1-2 h.

[0023] Preferably, in step 6), the molar volume ratio of the acridine intermediate, N,N-dimethyl-p-phenylenediamine and phenol is 1 mmol: 3-5 mmol: 2-4 mL; the temperature of the nucleophilic substitution reaction is 90-110° C., and the time of the nucleophilic substitution reaction is 1-3 h.

[0024] Preferably, in step 7), the molar volume ratio of the trisubstituted acridine intermediate 1 to sulfuric acid is 2-4 mmol:10 mL, and the mass concentration of sulfuric acid is 92-98%; the temperature of the hydrolysis reaction is 90-110° C., and the time of the hydrolysis reaction is 20-40 min;

[0025] Step 8) The molar volume ratio of the trisubstituted acridine intermediate 2,3-chloropropionyl chloride, triethylamine and N,N-dimethylformamide is 1mmol:4~6mmol:0.3~0.6mL:1~3mL; the temperature of the nucleophilic addition reaction is 50~70°C, and the time of the nucleophilic addition reaction is 1~3h.

[0026] The present invention also provides the use of the trisubstituted acridine derivative in the preparation of anti-tumor drugs.

[0027] Beneficial effects of the present invention:

[0028] The trisubstituted acridine derivatives of the present invention have excellent selectivity and stability for quadruple helix structure (G4-DNA), and have a broad spectrum of anti-tumor activity, including but not limited to ovarian cancer, prostate cancer, colorectal cancer, lung cancer, liver cancer, gastric cancer, etc., and have low toxic side effects on normal cells and tissues. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The trisubstituted acridine derivative prepared in Example 1 1 H NMR nuclear magnetic resonance spectrum;

[0030] Figure 2 The changes of body weight, heart weight and liver weight of mice in biological group and control group;

[0031] Figure 3 The changes of spleen weight, lung weight and kidney weight of mice in biological group and control group;

[0032] Figure 4 is the content of ALT, AST and Cr in the serum of mice in the biological group and the control group;

[0033] Figure 5 This is the selective detection result of the trisubstituted acridine derivative prepared in Example 1 to G4-DNA. DETAILED DESCRIPTION

[0034] The present invention provides a tri-substituted acridine derivative, the structural formula of the tri-substituted acridine derivative is:

[0035] .

[0036] The present invention also provides a method for preparing the trisubstituted acridine derivative, comprising the following steps:

[0037] 1) condensing 2-chloro-4-aminobenzoic acid and acetic anhydride to obtain a condensation intermediate;

[0038] 2) Mix the condensation intermediate, m-aminoacetanilide, copper powder, cuprous oxide, potassium carbonate and ethylene glycol ether, and perform Ullmann reaction to obtain a benzoic acid intermediate;

[0039] 3) The benzoic acid intermediate and sulfuric acid are subjected to a cyclization reaction to obtain the acridone intermediate 1;

[0040] 4) subjecting the acridone intermediate 1 to a condensation reaction with butyric anhydride to obtain the acridone intermediate 2;

[0041] 5) The acridone intermediate 2 is subjected to an electrophilic substitution reaction with phosphorus oxychloride to obtain an acridine intermediate;

[0042] 6) Mixing the acridine intermediate, N,N-dimethyl-p-phenylenediamine and phenol, and performing a nucleophilic substitution reaction to obtain a trisubstituted acridine intermediate 1;

[0043] 7) hydrolyzing the trisubstituted acridine intermediate 1 with sulfuric acid to obtain the trisubstituted acridine intermediate 2;

[0044] 8) The trisubstituted acridine intermediate 2,3-chloropropionyl chloride, triethylamine and N,N-dimethylformamide are mixed and subjected to a nucleophilic addition reaction to obtain a trisubstituted acridine derivative.

[0045] In the present invention, the synthesis route of the trisubstituted acridine derivative is preferably:

[0046] .

[0047] In the present invention, the molar volume ratio of 2-chloro-4-aminobenzoic acid and acetic anhydride in step 1) is preferably 110-120 mmol:30 mL, more preferably 112-118 mmol:30 mL, and more preferably 116.6 mmol:30 mL; the temperature of the condensation reaction is preferably 90-110° C., more preferably 95-105° C., and more preferably 100° C.; the condensation reaction time is preferably 20-40 min, more preferably 25-35 min, and more preferably 30 min.

[0048] In the present invention, the condensation reaction in step 1) is preferably stirred, and the stirring speed is preferably 700-900 rpm, and more preferably 800 rpm.

[0049] In the present invention, after the condensation reaction in step 1) is completed, dichloromethane is preferably added to precipitate, and then filtered and washed to obtain a condensation intermediate; the volume ratio of dichloromethane to acetic anhydride is preferably 2-4:1, more preferably 3:1; the reagent used for washing is preferably dichloromethane, and the number of washings is preferably 2-4 times, more preferably 3 times. The purpose of filtering is to remove excess acetic anhydride, by-product acetic acid and dichloromethane.

[0050] In the present invention, the condensation intermediate in step 1) is 4-(acetylamino)-2-chlorobenzoic acid, and the structural formula is: .

[0051] In the present invention, the molar ratio of the condensation intermediate, m-aminoacetanilide, copper powder, cuprous oxide and potassium carbonate in step 2) is preferably 1: 0.8-1.2: 0.08-0.12: 0.08-0.12: 0.2-0.8, more preferably 1: 0.9-1.1: 0.09-0.11: 0.09-0.11: 0.3-0.6, more preferably 1: 1.0: 0.098: 0.092: 0.5; the molar volume ratio of the condensation intermediate and ethylene glycol ethyl ether is preferably 1 mmol: 1-2.5 mL, more preferably 1 mmol: 1.3-2 mL, more preferably 1 mmol: 1.7 mL; the temperature of the Ullmann reaction is preferably 125-145 ° C, more preferably 130-140 ° C, more preferably 135 ° C; the time of the Ullmann reaction is preferably 4-6 h, more preferably 4.5-5.5 h, more preferably 5 h.

[0052] In the present invention, stirring is preferably performed during the Ullmann reaction in step 2), and the stirring speed is preferably 400-600 rpm, and more preferably 500 rpm.

[0053] In the present invention, after the Ullmann reaction in step 2) is completed, it is preferred to sequentially perform quenching reaction, filtration, recrystallization, and drying to obtain a benzoic acid intermediate; the reagent used in the quenching reaction is water, and the volume ratio of water to ethylene glycol ether is preferably 3-4:1, and more preferably 3.75:1; the recrystallization is preferably acidified by using hydrochloric acid to acidify the filtrate obtained by filtration, and the volume ratio of HCl to water in the hydrochloric acid is preferably 2.5-3.5:1, and more preferably 3:1, and the acidification is completed when the pH value of the filtrate is 2-3; the drying temperature is preferably 40-60°C, and more preferably 50°C; the drying time is preferably 20-28h, and more preferably 24h. The purpose of filtration is to remove the catalyst copper powder and cuprous oxide.

[0054] In the present invention, the benzoic acid intermediate in step 2) is 4-(acetylamino)-2-(3-(acetylamino)phenylamino)benzoic acid, and the structural formula is: .

[0055] In the present invention, the molar volume ratio of the benzoic acid intermediate and sulfuric acid in step 3) is preferably 14-17 mmol:12 mL, more preferably 15-16 mmol:12 mL, and more preferably 15.3 mmol:12 mL; the mass concentration of sulfuric acid is preferably 92-98%, and more preferably 94-96%; the temperature of the cyclization reaction is preferably 125-145°C, and more preferably 130-140°C, and more preferably 135°C; the time of the cyclization reaction is preferably 2-4 h, and more preferably 2.5-3.5 h, and more preferably 3 h.

[0056] In the present invention, the cyclization reaction in step 3) is preferably stirred, and the stirring speed is preferably 400-600 rpm, and more preferably 500 rpm.

[0057] In the present invention, after the cyclization reaction in step 3) is completed, the reaction solution is preferably placed in ice water to obtain a crude mixture, and the crude mixture is sequentially recrystallized, filtered, and dried to obtain an acridone intermediate 1; the temperature of the ice water is preferably -10~10°C, more preferably 0°C, and the volume ratio of ice water to sulfuric acid is preferably 100:10~15, more preferably 100:12; the recrystallization is preferably performed by adjusting the pH value of the crude mixture to 12~13 using a saturated sodium hydroxide solution; the drying temperature is preferably 40~60°C, more preferably 50°C; the drying time is preferably 20~28h, more preferably 24h.

[0058] In the present invention, the acridone intermediate 1 in step 3) is 3,6-diamino-9(10H)-acridone, and the structural formula is: .

[0059] In the present invention, the molar volume ratio of the acridone intermediate 1 and butyric anhydride in step 4) is preferably 0.8~2.5mmol:10mL, more preferably 1~2mmol:10mL, and more preferably 1.33mmol:10mL; the temperature of the condensation reaction is preferably 90~110°C, more preferably 95~105°C, and more preferably 100°C; the condensation reaction time is preferably 1~3h, more preferably 1.5~2.5h, and more preferably 2h.

[0060] In the present invention, the condensation reaction in step 4) is preferably stirred, and the stirring speed is preferably 400-600 rpm, and more preferably 500 rpm.

[0061] In the present invention, after the condensation reaction in step 4) is completed, recrystallization, filtration and drying are preferably performed in sequence to obtain the acridone intermediate 2; the recrystallization is preferably performed by adjusting the pH value of the solution after the reaction to 12-13 using a sodium hydroxide solution, and the mass concentration of the sodium hydroxide solution is preferably 37-43%, more preferably 40%; the drying temperature is preferably 40-60°C, more preferably 50°C; the drying time is preferably 20-28h, more preferably 24h.

[0062] In the present invention, the acridone intermediate 2 in step 4) is 3,6-bis(butyrylamino)-9(10H)-acridone, and the structural formula is: .

[0063] In the present invention, the molar volume ratio of the acridone intermediate 2 and phosphorus oxychloride in step 5) is preferably 12-15 mmol:20 mL, more preferably 13-14 mmol:20 mL, and more preferably 13.7 mmol:20 mL; the temperature of the electrophilic substitution reaction is preferably 80-100° C., more preferably 85-95° C., and more preferably 90° C.; the time of the electrophilic substitution reaction is preferably 1-2 h, and more preferably 1.5 h.

[0064] In the present invention, the electrophilic substitution reaction in step 5) is preferably stirred, and the stirring speed is preferably 400-600 rpm, and more preferably 500 rpm.

[0065] In the present invention, after the electrophilic substitution reaction in step 5) is completed, the reaction solution is preferably cooled to room temperature, and vacuum distillation, neutralization of phosphorus oxychloride, and extraction are sequentially performed to obtain an organic layer, and the organic layer is sequentially dried, vacuum distilled, and silica gel column chromatography is performed to obtain an acridine intermediate; the vacuum distillation temperature is independently preferably 50-60°C, and more preferably 55°C; the vacuum distillation pressure is independently preferably 0.05-0.15MPa, and more preferably 0.1MPa; the vacuum distillation time is independently preferably 5-15min, and more preferably 10min; the neutralization of phosphorus oxychloride is preferably to place the vacuum distillation product in ice water to obtain a crude product, and saturated sodium hydroxide is used to dissolve the crude product. The pH value of the crude product is adjusted to 12-13 by adding liquid; the temperature of the ice water is preferably -10-10°C, more preferably 0°C, the volume ratio of ice water to phosphorus oxychloride is preferably 100:15-25, more preferably 100:20; the reagents used in the extraction are preferably dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is preferably 9-11:1, more preferably 10:1; the number of extractions is preferably 2-4 times, more preferably 3 times; the reagent used in the drying is preferably anhydrous magnesium sulfate; the reagents used in the silica gel column chromatography are preferably dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is preferably 20-40:1, more preferably 30:1. The purpose of the first vacuum distillation is to remove the solvent phosphorus oxychloride, and the purpose of the second vacuum distillation is to remove dichloromethane and methanol.

[0066] In the present invention, the acridine intermediate in step 5) is 3,6-bis(butyrylamino)-9-chloroacridine, and the structural formula is: .

[0067] In the present invention, the molar volume ratio of the acridine intermediate, N,N-dimethyl-p-phenylenediamine and phenol in step 6) is preferably 1mmol:3~5mmol:2~4mL, more preferably 1mmol:3.5~4.5mmol:2.5~3.5mL, more preferably 1mmol:4mmol:2.88mL; the temperature of the nucleophilic substitution reaction is preferably 90~110°C, more preferably 95~105°C, more preferably 100°C; the time of the nucleophilic substitution reaction is preferably 1~3h, more preferably 1.5~2.5h, more preferably 2h.

[0068] In the present invention, the nucleophilic substitution reaction in step 6) is preferably stirred, and the stirring speed is preferably 400-600 rpm, and more preferably 500 rpm.

[0069] In the present invention, after the nucleophilic substitution reaction in step 6) is completed, it is preferred to sequentially perform a quenching reaction and an extraction to obtain an organic layer, and the organic layer is sequentially dried, distilled under reduced pressure, and subjected to silica gel column chromatography to obtain a trisubstituted acridine intermediate 1; the reagent used in the quenching reaction is preferably water, and the volume ratio of water to phenol is preferably 100:10-15, and more preferably 100:12; the reagent used in the extraction is preferably dichloromethane and a saturated sodium chloride solution, and the volume ratio of dichloromethane and a saturated sodium chloride solution is preferably 1-2:1, and more preferably 1.5:1; the extraction is repeated for a period of time. The number of times is preferably 2 to 4 times, more preferably 3 times; the reagent used for the drying is preferably anhydrous magnesium sulfate; the temperature of the vacuum distillation is preferably 50 to 60°C, more preferably 55°C; the pressure of the vacuum distillation is preferably 0.05 to 0.15 MPa, more preferably 0.1 MPa; the time of the vacuum distillation is preferably 10 to 20 minutes, more preferably 15 minutes; the reagents used for the silica gel column chromatography are preferably dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is preferably 10 to 30:1, more preferably 20:1. The purpose of the vacuum distillation is to remove dichloromethane and methanol.

[0070] In the present invention, the trisubstituted acridine intermediate 1 in step 6) is 3,6-bis(butyrylamino)-9-[4'-(N,N-dimethylamino)phenylamino]acridine, and the structural formula is: .

[0071] In the present invention, the molar volume ratio of the trisubstituted acridine intermediate 1 and sulfuric acid in step 7) is preferably 2-4 mmol:10 mL, more preferably 2.68-3.5 mmol:10 mL, and more preferably 3 mmol:10 mL; the mass concentration of sulfuric acid is preferably 92-98%, and more preferably 94-96%; the temperature of the hydrolysis reaction is preferably 90-110° C., and more preferably 95-105° C., and more preferably 100° C.; the time of the hydrolysis reaction is preferably 20-40 min, and more preferably 25-35 min, and more preferably 30 min.

[0072] In the present invention, the hydrolysis reaction in step 7) is preferably stirred, and the stirring speed is preferably 400-600 rpm, and more preferably 500 rpm.

[0073] In the present invention, after the hydrolysis reaction in step 7) is completed, the reaction solution is preferably cooled to room temperature, and sulfuric acid neutralization, recrystallization, filtration, and drying are sequentially performed to obtain the trisubstituted acridine intermediate 2; the neutralization of sulfuric acid is preferably performed by placing the reaction solution in ice water to obtain a crude mixture; the temperature of the ice water is preferably -10~10°C, more preferably 0°C, and the volume ratio of ice water to sulfuric acid is preferably 50:8~12, more preferably 50:10; the recrystallization is preferably performed by adjusting the pH value of the crude mixture to 12~13 using a saturated sodium hydroxide solution; the drying temperature is preferably 40~50°C, more preferably 45°C; the drying time is preferably 20~28h, more preferably 24h.

[0074] In the present invention, the trisubstituted acridine intermediate 2 in step 7) is 3,6-diamino-9-[4'-(N,N-dimethylamino)phenylamino]acridine, and the structural formula is: .

[0075] Step 8) The molar volume ratio of the trisubstituted acridine intermediate 2,3-chloropropionyl chloride, triethylamine and N,N-dimethylformamide is preferably 1mmol:4~6mmol:0.3~0.6mL:1~3mL, more preferably 1mmol:4.5~5.5mmol:0.4~0.5mL:1.67~2.5mL, more preferably 1mmol:5mmol:0.42mL:2mL; the temperature of the nucleophilic addition reaction is preferably 50~70°C, more preferably 55~65°C, more preferably 60°C; the time of the nucleophilic addition reaction is preferably 1~3h, more preferably 1.5~2.5h, more preferably 2h.

[0076] In the present invention, the nucleophilic addition reaction in step 8) is preferably stirred, and the stirring speed is preferably 400-600 rpm, and more preferably 500 rpm.

[0077] In the present invention, in step 8), the mixing preferably comprises first mixing the trisubstituted acridine intermediate 2 and triethylamine, and then sequentially adding N,N-dimethylformamide and 3-chloropropionyl chloride.

[0078] In the present invention, after the nucleophilic addition reaction in step 8) is completed, a quenching reaction, a reduced pressure distillation, and an extraction are sequentially performed to obtain an organic layer, and the organic layer is sequentially dried, distilled under reduced pressure, and subjected to silica gel column chromatography to obtain a trisubstituted acridine derivative; the reagent used in the quenching reaction is water, and the volume ratio of water to N,N-dimethylformamide is preferably 20-30:1, and more preferably 25:1; the temperature of the reduced pressure distillation is independently preferably 60-70°C, and more preferably 65°C; the pressure of the reduced pressure distillation is independently preferably 0.05-0.15MPa, and more preferably 0.1MP a; the time of reduced pressure distillation is independently preferably 20-30 min, more preferably 25 min; the reagents used for the extraction are preferably dichloromethane and saturated sodium chloride solution, and the volume ratio of dichloromethane to saturated sodium chloride solution is preferably 1-2:1, more preferably 1.5; 1; the number of extractions is preferably 2-4 times, more preferably 3 times; the reagent used for drying is preferably anhydrous magnesium sulfate; the reagents used for the silica gel column chromatography are preferably dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is preferably 10-30:1, more preferably 20:1.

[0079] In the present invention, the trisubstituted acridine derivative in step 8) is N,N'-(9-(4-(dimethylamino)phenylamino)acridine-3,6-diyl)bis(3-chloropropionamide).

[0080] The present invention also provides the use of the trisubstituted acridine derivative in the preparation of anti-tumor drugs.

[0081] In the present invention, the anti-tumor drug is preferably an injection, tablet, capsule, aerosol, suppository, film, pill, ointment, controlled release agent, sustained release agent or nano preparation.

[0082] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0083] Example 1

[0084] 116.6mmol 2-chloro-4-aminobenzoic acid and 30mL acetic anhydride were mixed and stirred at 100°C at 800rpm for 30min for condensation reaction to obtain a clear solution. 90mL dichloromethane was added to the clear solution to precipitate, and then filtered with a sand core funnel to obtain a solid product. The solid product was washed with dichloromethane 3 times to obtain a condensation intermediate: 4-(acetylamino)-2-chlorobenzoic acid. The condensation intermediate was a white solid with a mass of 17.2g and a yield of 69.1%.

[0085] 2.34mmol of anhydrous potassium carbonate, 0.46mmol of copper powder, 0.43mmol of cuprous oxide, 4.68mmol of condensation intermediate, 5.15mmol of m-aminoacetanilide and 8mL of ethylene glycol ether were mixed and stirred at 135°C at a speed of 500rpm for 5h to perform Ullmann reaction to obtain a reaction mixture. The reaction mixture was poured into 30mL of distilled water and filtered with a diatomaceous earth filter to obtain a filtrate. Hydrochloric acid (the volume ratio of HCl to water in hydrochloric acid is 3:1) was added to the filtrate for acidification, and the acidification was terminated when the pH value of the filtrate was 2.5, and the crude product was filtered again to obtain a crude product. The crude product was placed at 50°C and dried for 24h to obtain a benzoic acid intermediate: 4-(acetylamino)-2-(3-(acetylamino)phenylamino)benzoic acid. The benzoic acid intermediate is a gray solid with a mass of 420mg and a yield of 27.4%.

[0086] 12 mL of 96% sulfuric acid was added to 15.3 mmol of benzoic acid intermediate at a rate of 1 drop / s, and the mixture was stirred at 135°C and 500 rpm for 2 h for cyclization reaction. After the reaction, the mixture was cooled to room temperature to obtain a reaction mixture. The reaction mixture was poured into 100 mL of ice water at 0°C to obtain a crude mixture, and the pH value of the crude mixture was adjusted to 12.5 with a saturated sodium hydroxide solution, and then filtered with a sand core funnel to obtain a filter residue, which was dried at 50°C for 24 h to obtain acridone intermediate 1: 3,6-diamino-9(10H)-acridone. Acridone intermediate 1 was a yellow-green solid and was directly used in the next reaction step without further purification.

[0087] 1.33mmol of acridone intermediate 1 was placed in a 100mL round-bottom flask, 10mL of butyric anhydride was added, and the mixture was stirred at 100°C and 500rpm for 2h for condensation reaction to obtain a reaction mixture. The pH value of the reaction mixture was adjusted to 12.5 with a sodium hydroxide solution having a mass concentration of 40%, and then filtered with a sand core funnel to obtain a filter residue, which was placed at 50°C and dried for 24h to obtain acridone intermediate 2: 3,6-bis(butyrylamino)-9(10H)-acridone. Acridone intermediate 2 was a gray solid and was directly used in the next reaction step without further purification.

[0088] Add 20 mL of phosphorus oxychloride to 13.7 mmol of acridone intermediate 2, stir at 90°C at 500 rpm for 1.5 h to perform an electrophilic substitution reaction, and cool to room temperature after the reaction to obtain a reaction mixture. Place the reaction mixture in a rotary evaporator, distill under reduced pressure at 55°C and 0.1 MPa for 10 min, pour the product of reduced pressure distillation into 100 mL of ice water at 0°C to obtain a crude product, and adjust the pH value of the crude product to 12.5 with a saturated sodium hydroxide solution to obtain a mixed solution. Use dichloromethane and methanol with a volume ratio of 10:1 to extract the mixed solution, extract 3 times to obtain an organic layer, dry the organic layer with anhydrous magnesium sulfate, place it in a rotary evaporator, and distill under reduced pressure at 55°C and 0.1 MPa for 15 min. The product of vacuum distillation was subjected to silica gel column chromatography using dichloromethane and methanol in a volume ratio of 30:1 (the drop rate of the product of vacuum distillation was 5 drops / s) to obtain an acridine intermediate: 3,6-bis(butyrylamino)-9-chloroacridine. The acridine intermediate was a bright yellow solid with a mass of 1.6 g and a yield of 30.5%.

[0089] 4.16mmol of acridine intermediate, 16.66mmol of N,N-dimethyl-p-phenylenediamine and 12mL of phenol were mixed, stirred at 100℃ and 500rpm for 2h to perform nucleophilic substitution reaction to obtain a reaction mixture. The reaction mixture was poured into 100mL of distilled water, the pH value of the reaction mixture was adjusted to 12.5 with saturated sodium hydroxide solution, and then extracted three times with dichloromethane and saturated sodium chloride solution in a volume ratio of 1:1 to obtain an organic layer, which was dried with anhydrous magnesium sulfate and placed in a rotary evaporator, and distilled under reduced pressure at 55℃ and 0.1MPa for 15min. The product of reduced pressure distillation was subjected to silica gel column chromatography with dichloromethane and methanol in a volume ratio of 20:1 (the drop rate of the product of reduced pressure distillation was 5 drops / s) to obtain a trisubstituted acridine intermediate 1:3,6-bis(butyrylamino)-9-[4'-(N,N-dimethylamino)phenylamino]acridine. The trisubstituted acridine intermediate 1 is a reddish brown solid with a mass of 1.3 g and a yield of 64.7%.

[0090] 10 mL of 96% sulfuric acid was added to 2.68 mmol of trisubstituted acridine intermediate 1 at a rate of 1 drop / s, and the mixture was stirred at 500 rpm for 0.5 h at 100 ° C for hydrolysis reaction. After the reaction, the mixture was cooled to room temperature to obtain a reaction mixture. The reaction mixture was poured into 50 mL of ice water at 0 ° C to obtain a crude product, and the pH value of the crude product was adjusted to 12.5 with a saturated sodium hydroxide solution, and then filtered with a sand core funnel to obtain a filter residue, and the filter residue was placed at 45 ° C and dried for 24 h to obtain a trisubstituted acridine intermediate 2: 3,6-diamino-9-[4'-(N,N-dimethylamino)phenylamino]acridine. The trisubstituted acridine intermediate 2 is a reddish brown solid, which can be directly used in the next reaction step without further purification.

[0091] 2.39mmol of trisubstituted acridine intermediate 2 and 1mL of triethylamine were mixed, and then 4mL of N,N-dimethylformamide and 11.95mmol of 3-chloropropionyl chloride were added in sequence, and the mixture was stirred at 60°C and 500rpm for 2h for nucleophilic addition reaction to obtain a reaction mixture. The reaction mixture was poured into 100mL of distilled water and placed in a rotary evaporator for vacuum distillation at 65°C and 0.1MPa for 25min. The product of vacuum distillation was extracted three times with dichloromethane and saturated sodium chloride solution in a volume ratio of 1:1 to obtain an organic layer. The organic layer was dried with anhydrous magnesium sulfate and placed in a rotary evaporator for vacuum distillation at 60°C and 0.1MPa for 20min. The product of the vacuum distillation was subjected to silica gel column chromatography using dichloromethane and methanol in a volume ratio of 20:1 (the dropping rate of the product of the vacuum distillation was 5 drops / s) to obtain a trisubstituted acridine derivative: N,N'-(9-(4-(dimethylamino)phenylamino)acridine-3,6-diyl)bis(3-chloropropionamide). The trisubstituted acridine derivative was a reddish brown solid with a mass of 550 mg and a yield of 44.7%.

[0092] Example 2

[0093] 110 mmol of 2-chloro-4-aminobenzoic acid and 30 mL of acetic anhydride were mixed and stirred at 110° C. and 800 rpm for 20 min to perform a condensation reaction to obtain a clear solution. 120 mL of dichloromethane was added to the clear solution to precipitate a precipitate, which was then filtered using a sand core funnel to obtain a solid product, which was washed three times with dichloromethane to obtain a condensation intermediate: 4-(acetylamino)-2-chlorobenzoic acid.

[0094] 3.74mmol of anhydrous potassium carbonate, 0.56mmol of copper powder, 0.56mmol of cuprous oxide, 4.68mmol of condensation intermediate, 3.75mmol of m-aminoacetanilide and 9.5mL of ethylene glycol ether were mixed and stirred at 145°C for 4h at a speed of 600rpm for Ullmann reaction to obtain a reaction mixture. The reaction mixture was poured into 38mL of distilled water and filtered with a diatomaceous earth filter to obtain a filtrate. Hydrochloric acid (the volume ratio of HCl to water in hydrochloric acid is 3:1) was added to the filtrate for acidification, and the acidification was terminated when the pH value of the filtrate was 2.5, and the crude product was filtered again. The crude product was placed at 40°C and dried for 28h to obtain a benzoic acid intermediate: 4-(acetylamino)-2-(3-(acetylamino)phenylamino)benzoic acid.

[0095] 12 mL of 92% sulfuric acid was added to 17 mmol of benzoic acid intermediate at a rate of 1 drop / s, and the mixture was stirred at 400 rpm for 4 h at 125 ° C for cyclization reaction. After the reaction, the mixture was cooled to room temperature to obtain a reaction mixture. The reaction mixture was poured into 80 mL of ice water at 0 ° C to obtain a crude mixture, and the pH value of the crude mixture was adjusted to 12 with a saturated sodium hydroxide solution, and then filtered with a sand core funnel to obtain a filter residue, which was placed at 40 ° C and dried for 28 h to obtain acridone intermediate 1: 3,6-diamino-9 (10H) -acridone. Acridone intermediate 1 is a yellow-green solid and is directly used in the next reaction step without further purification.

[0096] 2mmol of acridone intermediate 1 was placed in a 100mL round-bottom flask, 10mL of butyric anhydride was added, and the mixture was stirred at 90°C for 3h at a speed of 400rmp for condensation reaction to obtain a reaction mixture. The pH value of the reaction mixture was adjusted to 13 with a sodium hydroxide solution having a mass concentration of 43%, and then filtered with a sand core funnel to obtain a filter residue, which was placed at 60°C and dried for 20h to obtain acridone intermediate 2: 3,6-bis(butyrylamino)-9(10H)-acridone. Acridone intermediate 2 was a gray solid and was directly used in the next reaction step without further purification.

[0097] Add 20 mL of phosphorus oxychloride to 15 mmol of acridone intermediate 2, stir at 100 ° C at 600 rpm for 1 h to carry out electrophilic substitution reaction, and cool to room temperature after the reaction to obtain a reaction mixture. The reaction mixture was placed in a rotary evaporator and distilled under reduced pressure at 50 ° C and 0.15 MPa for 15 min. The product of the reduced pressure distillation was poured into 80 mL of ice water at 0 ° C to obtain a crude product, and the pH value of the crude product was adjusted to 12 with a saturated sodium hydroxide solution to obtain a mixed solution. The mixed solution was extracted with dichloromethane and methanol in a volume ratio of 9:1, and the organic layer was obtained by extraction 3 times. The organic layer was dried with anhydrous magnesium sulfate and placed in a rotary evaporator, and distilled under reduced pressure at 50 ° C and 0.15 MPa for 15 min. The product of vacuum distillation was subjected to silica gel column chromatography using dichloromethane and methanol in a volume ratio of 40:1 (the drop rate of the product of vacuum distillation was 5 drops / s) to obtain an acridine intermediate: 3,6-bis(butyrylamino)-9-chloroacridine. The acridine intermediate was a bright yellow solid with a mass of 1.6 g and a yield of 30.5%.

[0098] 4.16mmol of acridine intermediate, 20mmol of N,N-dimethyl-p-phenylenediamine and 16.5mL of phenol were mixed, stirred at 90℃ and 400rpm for 3h to perform nucleophilic substitution reaction to obtain a reaction mixture. The reaction mixture was poured into 110mL of distilled water, the pH value of the reaction mixture was adjusted to 12 with saturated sodium hydroxide solution, and then extracted three times with dichloromethane and saturated sodium chloride solution in a volume ratio of 2:1 to obtain an organic layer, which was dried with anhydrous magnesium sulfate and placed in a rotary evaporator, and distilled under reduced pressure at 50℃ and 0.15MPa for 10min. The product of reduced pressure distillation was subjected to silica gel column chromatography with dichloromethane and methanol in a volume ratio of 10:1 (the drop rate of the product of reduced pressure distillation was 5 drops / s) to obtain a trisubstituted acridine intermediate 1:3,6-bis(butyrylamino)-9-[4'-(N,N-dimethylamino)phenylamino]acridine. The trisubstituted acridine intermediate 1 is a reddish brown solid with a mass of 1.3 g and a yield of 64.7%.

[0099] 10 mL of 92% sulfuric acid was added to 4 mmol of trisubstituted acridine intermediate 1 at a rate of 1 drop / s, and the mixture was stirred at 400 rpm for 40 min at 90°C for hydrolysis reaction. After the reaction, the mixture was cooled to room temperature to obtain a reaction mixture. The reaction mixture was poured into 42 mL of ice water at 0°C to obtain a crude product, and the pH value of the crude product was adjusted to 13 with a saturated sodium hydroxide solution, and then filtered with a sand core funnel to obtain a filter residue, which was placed at 40°C and dried for 28 hours to obtain a trisubstituted acridine intermediate 2: 3,6-diamino-9-[40-(N,N-dimethylamino)phenylamino]acridine. The trisubstituted acridine intermediate 2 was a reddish brown solid and was directly used in the next reaction step without further purification.

[0100] 2.39mmol of trisubstituted acridine intermediate 2 and 1.4mL of triethylamine were mixed, and then 7mL of N,N-dimethylformamide and 14mmol of 3-chloropropionyl chloride were added in sequence, and the mixture was stirred at 70°C and 500rpm for 2h for nucleophilic addition reaction to obtain a reaction mixture. The reaction mixture was poured into 140mL of distilled water, and placed in a rotary evaporator, and distilled under reduced pressure at 60°C and 0.05MPa for 20min. The product of the reduced pressure distillation was extracted three times with dichloromethane and saturated sodium chloride solution in a volume ratio of 1:1 to obtain an organic layer. The organic layer was dried with anhydrous magnesium sulfate and placed in a rotary evaporator, and distilled under reduced pressure at 60°C and 0.1MPa for 15min. The product of the vacuum distillation was subjected to silica gel column chromatography using dichloromethane and methanol in a volume ratio of 30:1 (the dropping rate of the vacuum distillation product was 5 drops / s) to obtain a trisubstituted acridine derivative: N,N'-(9-(4-(dimethylamino)phenylamino)acridine-3,6-diyl)bis(3-chloropropionamide).

[0101] The trisubstituted acridine derivative prepared in Example 1 was characterized by NMR. Figure 1 The trisubstituted acridine derivative prepared in Example 1 1 H NMR nuclear magnetic resonance spectrum.

[0102] Figure 1 Income 1 H NMR is: 1 H NMR (400 MHz, DMSO- d 6) δ 10.93 (s, 2H), 8.48 (s,2H), 8.11 (d, J = 9.3 Hz, 2H), 7.41 – 7.18 (m, 4H), 6.84 (d, J = 8.5 Hz, 2H), 3.96 (s, 4H), 3.00 (s, 10H).

[0103] Depend on Figure 1 It can be seen that the trisubstituted acridine derivative prepared in Example 1 of the present invention is N,N'-(9-(4-(dimethylamino)phenylamino)acridine-3,6-diyl)bis(3-chloropropionamide).

[0104] The antitumor activity of the trisubstituted acridine derivative prepared in Example 1 was detected by MTT method, and the detection steps were as follows:

[0105] Ovarian cancer cells SKOV3 in the logarithmic growth phase were seeded in a 96-well plate (seeding density of 500 cells / well, 160 μL culture medium / well, the culture medium was high-glucose medium DMEM, the culture medium brand was GIBCO, the product number was C11965500BT, and the packaging specification was 500 mL), and the 96-well plate was placed in a 37°C incubator for 12 hours. The 96-well plate was taken out, and 40 μL of trisubstituted acridine derivative culture medium of different concentrations was added to different wells (the concentrations of trisubstituted acridine derivatives in the culture medium were 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM, 1.5625 μM, and 0.78125 μM, respectively, the culture medium was high-glucose medium DMEM, the culture medium brand was GIBCO, the product number was C11965500BT, and the packaging specification was 500 mL). Place the 96-well plate in a 37°C incubator and culture for 44 hours. Take out the 96-well plate, add 20 μL of 5 mg / mL MTT working solution (i.e., thiazolyl blue working solution, with normal saline as the solvent) to each well, and continue to culture in a 37°C incubator for 4 hours. After the culture is completed, use a vacuum pump to suck out the supernatant until there is no liquid remaining in the well, add 100 μL of dimethyl sulfoxide to each well, and oscillate in the dark at room temperature for 15 seconds. Determine the absorbance of the solution in each well at 492 nm to obtain the cell viability, and use GraphPad Prism9 to plot the cell viability against the concentration of the trisubstituted acridine derivative to obtain the IC value of the trisubstituted acridine derivative for ovarian cancer cells SKOV3. 50 Values ​​are shown in Table 1.

[0106] The above detection steps were repeated on prostate cancer cells DU145, colorectal cancer cells HCT116, lung cancer cells A549, liver cancer cells Huh-7 and gastric cancer cells SGC in the logarithmic growth phase to obtain the IC values ​​of trisubstituted acridine derivatives for different tumor cells. 50 Values ​​are shown in Table 1.

[0107] Table 1 Antitumor activity of trisubstituted acridine derivatives

[0108]

[0109] As can be seen from Table 1, the trisubstituted acridine derivatives of the present invention have good antitumor activity against ovarian cancer cells SKOV3, prostate cancer cells DU145, colorectal cancer cells HCT116, lung cancer cells A549, liver cancer cells Huh-7 and gastric cancer cells SGC7901. 50 The values ​​were all less than 20 μM. This result shows that the trisubstituted acridine derivatives of the present invention have a broad spectrum of antitumor activity.

[0110] The toxic and side effects of the trisubstituted acridine derivatives prepared in Example 1 on normal cells and tissues were detected. The detection steps were as follows:

[0111] Twenty ICR mice weighing 25g and aged 6 weeks were randomly divided into two groups (10 mice in each group), namely the biological group and the control group. The biological group was orally administered with a trisubstituted acridine derivative (the trisubstituted acridine derivative was dissolved in 100mL of normal saline) at a dose of 90mg / kg, and the control group was administered with the same dose of normal saline in the same way, and the body weight was recorded daily. After 7 days, the weights of the heart, liver, spleen, lung and kidney of the mice in the biological group and the control group were recorded respectively; and the eyeballs of the mice in the biological group and the control group were blood drawn, and the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and creatinine (Cr) in the serum were detected using alanine aminotransferase kit, aspartate aminotransferase kit and creatinine kit, respectively.

[0112] Figure 2 The changes of body weight, heart weight and liver weight of mice in biological group and control group are shown in Figure 2. Figure 3 The changes of spleen weight, lung weight and kidney weight of mice in biological group and control group are shown in Figure 2. Figure 4 is the content of ALT, AST and Cr in the serum of mice in the biological group and the control group.

[0113] Depend on Figures 2~4 It can be seen that compared with the control group, the administration of the trisubstituted acridine derivatives of the present invention does not significantly affect the weight of mice and the weight of each organ, nor does it significantly change the levels of ALT, AST and Cr in serum. This result shows that the trisubstituted acridine derivatives of the present invention have less damage and toxic side effects on normal cells and tissues and have higher biosafety.

[0114] The selectivity and stability of the trisubstituted acridine derivatives prepared in Example 1 to G4-DNA were tested by fluorescence resonance energy transfer competition experiment. The testing steps were as follows:

[0115] The human telomeric repeat sequence F21T: 5'-FAM-d(GGG[TTAGGG]3)-TAMRA-3' was used to artificially synthesize double-labeled G4-DNA, the fluorescent donor FAM was 6-carboxyfluorescein, and the fluorescent acceptor TAMRA was 6-carboxy-tetramethylrhodamine. The double-labeled G4-DNA was diluted to 400nM with a potassium arsenate buffer having a pH value of 7.4 and a concentration of 60mM; the calf thymus DNA was diluted to 40µM with a potassium arsenate buffer having a pH value of 7.4 and a concentration of 60mM. The trisubstituted acridine derivative prepared in Example 1 was prepared into a 3mM aqueous solution. The double-labeled G4-DNA solution and the calf thymus DNA solution were mixed to obtain mixed solutions with a concentration ratio of double-labeled G4-DNA to calf thymus DNA (CT-DNA) of 1:0, 1:1, 1:5, 1:10, 1:20, 1:50, 1:100 and 1:200, respectively. An aqueous solution of trisubstituted acridine derivatives less than 3% of the total volume of the mixed solution was added to the mixed solution, and the mixed working solution was balanced for 1 hour. 20 μL of the mixed working solution was injected into the LightCycler capillary, and the LightCycler capillary was inserted into the Roche LightCycler 480II fluorescence quantitative PCR instrument to detect the fluorescence emission intensity at 530 nm (the fixed excitation wavelength was 470 nm, the temperature increase range was 37-99 °C, the temperature increase interval was 1 °C, and the sample was collected after 30 seconds of equilibration each time the temperature was increased).

[0116] Figure 5 The results of the selective detection of the trisubstituted acridine derivatives prepared in Example 1 on G4-DNA. Figure 5 It can be seen that in the competition experiment between G4-DNA and CT-DNA, even if the concentration of CT-DNA is 200 times that of G4-DNA, the fluorescence intensity of G4-DNA cannot be reduced. This result shows that the trisubstituted acridine derivatives of the present invention have high selectivity and stability for G4-DNA.

[0117] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A trisubstituted acridine derivative, characterized in that: The structural formula of the trisubstituted acridine derivative is: 。 2. The method for preparing a trisubstituted acridine derivative according to claim 1, characterized in that: The following steps are included: (1) subjecting 2-chloro-4-aminobenzoic acid and acetic anhydride to a condensation reaction to obtain a condensation intermediate; (2) mixing the condensation intermediate, m-aminoacetanilide, copper powder, cuprous oxide, potassium carbonate and ethylene glycol ether, and performing an Ullmann reaction to obtain a benzoic acid intermediate; (3) The benzoic acid intermediate and sulfuric acid are subjected to a cyclization reaction to obtain an acridone intermediate 1; (4) subjecting acridone intermediate 1 to a condensation reaction with butyric anhydride to obtain acridone intermediate 2; (5) The acridone intermediate 2 is subjected to an electrophilic substitution reaction with phosphorus oxychloride to obtain an acridine intermediate; (6) Mixing the acridine intermediate, N,N-dimethyl-p-phenylenediamine and phenol, and performing a nucleophilic substitution reaction to obtain a trisubstituted acridine intermediate 1; (7) hydrolyzing the trisubstituted acridine intermediate 1 with sulfuric acid to obtain the trisubstituted acridine intermediate 2; (8) The trisubstituted acridine intermediate 2,3-chloropropionyl chloride, triethylamine and N,N-dimethylformamide are mixed and subjected to a nucleophilic addition reaction to obtain a trisubstituted acridine derivative.

3. The method for preparing a trisubstituted acridine derivative according to claim 2, characterized in that: In step (1), the molar volume ratio of 2-chloro-4-aminobenzoic acid to acetic anhydride is 110-120 mmol:30 mL; the temperature of the condensation reaction is 90-110° C., and the time of the condensation reaction is 20-40 min.

4. The method for preparing a trisubstituted acridine derivative according to claim 2 or 3, characterized in that: In step (2), the molar ratio of the condensation intermediate, m-aminoacetanilide, copper powder, cuprous oxide and potassium carbonate is 1:0.8-1.2:0.08-0.12:0.08-0.12:0.2-0.8, and the molar volume ratio of the condensation intermediate and ethylene glycol ethyl ether is 1 mmol:1-2.5 mL; the temperature of the Ullmann reaction is 125-145° C., and the time of the Ullmann reaction is 4-6 h.

5. The method for preparing a trisubstituted acridine derivative according to claim 4, characterized in that: In step (3), the molar volume ratio of the benzoic acid intermediate and sulfuric acid is 14-17 mmol:12 mL, and the mass concentration of sulfuric acid is 92-98%; the temperature of the cyclization reaction is 125-145° C., and the time of the cyclization reaction is 2-4 h.

6. The method for preparing a trisubstituted acridine derivative according to claim 5, characterized in that: In step (4), the molar volume ratio of the acridone intermediate 1 to butyric anhydride is 0.8-2.5 mmol:10 mL; the temperature of the condensation reaction is 90-110° C., and the condensation reaction time is 1-3 h.

7. The method for preparing a trisubstituted acridine derivative according to claim 5 or 6, characterized in that: In step (5), the molar volume ratio of the acridone intermediate 2 to phosphorus oxychloride is 12-15 mmol:20 mL; the temperature of the electrophilic substitution reaction is 80-100° C., and the time of the electrophilic substitution reaction is 1-2 h.

8. The method for preparing a trisubstituted acridine derivative according to claim 7, characterized in that: In step (6), the molar volume ratio of the acridine intermediate, N,N-dimethyl-p-phenylenediamine and phenol is 1 mmol: 3-5 mmol: 2-4 mL; the temperature of the nucleophilic substitution reaction is 90-110° C., and the time of the nucleophilic substitution reaction is 1-3 h.

9. The method for preparing a trisubstituted acridine derivative according to claim 8, characterized in that: In step (7), the molar volume ratio of the trisubstituted acridine intermediate 1 to sulfuric acid is 2-4 mmol:10 mL, and the mass concentration of sulfuric acid is 92-98%; the temperature of the hydrolysis reaction is 90-110° C., and the time of the hydrolysis reaction is 20-40 min; In step (8), the molar volume ratio of the trisubstituted acridine intermediate 2,3-chloropropionyl chloride, triethylamine and N,N-dimethylformamide is 1 mmol: 4-6 mmol: 0.3-0.6 mL: 1-3 mL; the temperature of the nucleophilic addition reaction is 50-70° C., and the time of the nucleophilic addition reaction is 1-3 h.

10. Use of the trisubstituted acridine derivative according to claim 1 in the preparation of anti-tumor drugs, characterized in that: The tumor is ovarian cancer, prostate cancer, colorectal cancer, lung cancer, liver cancer or gastric cancer.

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