CN114409632A

CN114409632A - 2-substituted amino-3-benzimidazole substituted quinoline derivative and preparation method and application thereof

Info

Publication number: CN114409632A
Application number: CN202210163145.8A
Authority: CN
Inventors: 潘成学; 李昕蔚; 方淑君; 李映泽; 黄雪球; 韦敏; 苏桂发
Original assignee: Guangxi Normal University
Current assignee: Beijing Kundayu Technology Co ltd
Priority date: 2022-02-22
Filing date: 2022-02-22
Publication date: 2022-04-29
Anticipated expiration: 2042-02-22
Also published as: CN114409632B

Abstract

The invention relates to the technical field of medicines, and particularly discloses a 2-substituted amino-3-benzimidazole substituted quinoline derivative, which has a structural formula as follows:

in the formula, R¹Is hydrogen, fluorine, chlorine, bromine, methyl or methoxy, R is dimethylamino, piperidinyl or tetrahydropyrrolyl, R is²Is hydrogen or methyl. The 2-substituted amino-3-benzimidazole substituted quinoline derivative can be used as a topoisomerase I inhibitor, has strong activity of inhibiting tumor cell proliferation, has in-vivo anti-tumor effect equivalent to that of the existing first medicament adriamycin for clinically treating bladder cancer, has the characteristics of high efficiency and low toxicity, and can be used for treating bladder cancerLead compounds of novel topoisomerase I inhibitors and antitumor drugs are developed.

Description

2-substituted amino-3-benzimidazole substituted quinoline derivative and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to 2-substituted amino-3-benzimidazole substituted quinoline derivatives, and a preparation method and application thereof.

Background

DNA topoisomerase I (DNA Topo I) is an important biological enzyme and mainly regulates and controls the topological structure in the processes of DNA replication, transcription, gene expression and the like. Research shows that high expression of topoisomerase I often appears in tumor cell tissues, so that inhibition of activity of topoisomerase I can effectively inhibit rapid proliferation of tumor cells, therefore, topoisomerase is an important target for research of antitumor drugs, and a plurality of compounds with topoisomerase inhibitory activity are widely used clinically as antitumor drugs.

From a structural point of view, topoisomerase I inhibitors encompass both the camptothecin derivatives and the non-camptothecin derivatives. From the viewpoint of mechanism of action, drugs inhibiting topoisomerase I can be divided into two broad classes, topoisomerase I poisoning agents and topoisomerase I catalytic inhibitors, both of which inhibit the activity of enzymatic DNA unwinding. Wherein the poisoning agent prevents DNA from recovering and stabilizes the DNA-enzyme complex, we refer to the cleavable complex, by intercalating enzymes catalyzing the nicks of DNA breaks, thereby generating single-stranded DNA breaks. Inhibitors inhibit the catalytic activity of the enzyme by direct interaction with the enzyme or by forming a molecular complex with DNA. Camptothecin and its derivatives in natural products are representative of topoisomerase I poisoning agents, and luteolin can be used as enzyme inhibitor of topoisomerase I.

The inventor designs and synthesizes the combination of two advantageous structural units of a quinoline ring and a benzimidazole ring according to an advantageous structure combination strategy in drug molecule design on the basis of a topoisomerase I pharmacophore model constructed in the early stage of a subject group to obtain a new skeleton of a quinoline compound, and finds that part of the obtained target compound can be used as a strong topoisomerase I inhibitor, these compounds also showed strong activity of inhibiting tumor cell proliferation, and in the T24 bladder cancer transplantable tumor model test, the in vivo anti-tumor effect of the representative compound is equivalent to the anti-tumor effect of the first medicament adriamycin for clinically treating bladder cancer at present, and the weight of a mouse receiving the drug is hardly influenced, and organ HE staining shows that the drug has the characteristics of high efficiency and low toxicity, and the drug can be used as a lead compound for researching and developing novel topoisomerase I inhibitors and antitumor drugs.

Disclosure of Invention

The invention aims to provide 2-substituted amino-3-benzimidazole substituted quinoline derivatives and a preparation method thereof, wherein the 2-substituted amino-3-benzimidazole substituted quinoline derivatives can be used as a novel topoisomerase I inhibitor and show good anti-tumor effect in vitro and in vivo.

In order to achieve the purpose, the invention provides 2-substituted amino-3-benzimidazole substituted quinoline derivatives, which have the structural formula:

in the formula, R¹Is hydrogen, fluorine, chlorine, bromine, methyl or methoxy, R is dimethylamino, piperidinyl or tetrahydropyrrolyl, R is²Is hydrogen or methyl.

The preparation method of the 2-substituted amino-3-benzimidazole substituted quinoline derivative comprises the steps of dissolving the compound shown in the formula (II) in an organic solvent, adding the compound shown in the formula (III), and carrying out reflux reaction under the heating condition; after the reaction is finished, decompressing to remove the solvent, separating and purifying to obtain the target compound shown in the formula (I), namely the 2-substituted amino-3-benzimidazole substituted quinoline derivative;

wherein, the synthetic route of the 2-substituted amino-3-benzimidazole substituted quinoline derivative is as follows:

in the formula, R¹Is hydrogen, fluorine, chlorine, bromine, methyl or methoxy, R is dimethylaminoRadicals, piperidine radicals or tetrahydropyrrole radicals, R²Is hydrogen or methyl, and n is 2.

Preferably, in the above preparation method, the compound of formula (II): the molar ratio of the compound of the formula (III) is 1: 1.5-2.0.

Preferably, in the above preparation method, the reaction solvent may be ethanol or toluene.

Preferably, in the preparation method, the reaction temperature is reflux temperature, and the reaction time is 10-12 h.

Preferably, in the above preparation method, the heating reaction is performed under electromagnetic stirring conditions.

Preferably, in the above preparation method, the separation and purification is silica gel column chromatography separation and purification, gradient elution is performed, the eluent is dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is 100: 1.

The 2-substituted amino-3-benzimidazole substituted quinoline derivative is applied to the development of a novel topoisomerase I inhibitor.

The 2-substituted amino-3-benzimidazole substituted quinoline derivative is applied to preparing the antineoplastic drugs based on topoisomerase I.

Compared with the prior art, the invention has the following beneficial effects:

the 2-substituted amino-3-benzimidazole substituted quinoline derivative can be used as a topoisomerase I inhibitor, has strong activity of inhibiting tumor cell proliferation, has in-vivo anti-tumor effect equivalent to that of the existing first medicament adriamycin for clinically treating bladder cancer, has the characteristics of high efficiency and low toxicity, and can be used for developing novel topoisomerase I inhibitors and lead compounds of anti-tumor medicaments. The preparation method of the 2-substituted amino-3-benzimidazole substituted quinoline derivative has the advantages of simple operation, short preparation period, lower cost and stable quality.

Drawings

FIG. 1 is an experimental electrophoretogram of Topo I inhibitory activity in 100. mu.M of the compound of application example 1 of the present invention, lanes 1 and 19 (D): DNA, lanes 2 and 20 (T): DNA + Topo I, lanes 3 and 21 (C): CPT (100 μ M) + DNA + Topo I, lanes 4 and 22 (T): DNA + Topo I, lanes 5-18, 23-26: the selected cytotoxic compound (100 μ M each) + DNA + Topo I.

FIG. 2 is an electrophoretogram of relaxation experiment mediated by DNA topoisomerase I of application example 1 of the present invention, in (B, C, D): lane 1 (D): DNA, lane 2 (T): DNA + Topo I, lane 3 (C): CPT (100 μ M) + DNA + Topo I, lane 4 (M): luteolin (100 μ M) + DNA + Topo I, lanes 5-8: selected cytotoxic compound 2a (5, 10, 20, 50 μ M, respectively) + DNA + Topo I; (E, F, G) wherein: lanes 1-3: the cytotoxic compound 4c (10, 20, 50 μ M, respectively) + DNA + TopoI, lane 4 (D): DNA, lane 5 (T): DNA + Topo I, lane 6 (C): CPT (100 μ M) + DNA + Topo I, lane 7 (M): luteolin (300. mu.M) + DNA + Topo I.

FIG. 3 is an experimental electrophoretogram of Topo 1-DNA cleavable complex of Compound 2a in application example 2 of the present invention, in which lane 1 (D): DNA, lane 2 (T): DNA + Topo I, lane 3 (M): luteolin (300 μ M) + DNA + Topo I, lane 4 (S): DMSO (5%) + DNA + Topo I, lanes 5-7: selected cytotoxic compound 2a (5, 10, 20, 50 μ M) + DNA + Topo I, lane 8 (C): CPT (100. mu.M) + DNA + Topo I.

Fig. 4 is a comet image in application example 3 of the present invention.

FIG. 5 shows the effect of Compound 2a in application example 5 of the present invention on tumor weight and tumor inhibition rate of human bladder cancer cells T-24 nude mice xenografted with nude mice.

FIG. 6 is a photograph of each group of tumor-bearing nude mice and each group of tumor in application example 5 of the present invention.

FIG. 7 is a photograph showing HE staining of organ sections of tumor-bearing nude mice in each group in application example 5.

FIG. 8 is a graph showing the change in body weight of each group of tumor-bearing nude mice in application example 5 of the present invention.

Detailed Description

The following detailed description of specific embodiments of the invention is provided, but it should be understood that the scope of the invention is not limited to the specific embodiments.

The synthetic route of a series of target compounds 2-4a-h and 5-7 (2-substituted amino-3-benzimidazole substituted quinoline derivatives) is as follows:

the raw material compounds 1a-i are synthesized by the method provided in the Chinese invention patent with the publication number CN 105732576B.

Example 1

A2-substituted amino-3-benzimidazole substituted quinoline derivative 2a has a structural formula as follows:

the preparation method of the 2-substituted amino-3-benzimidazole substituted quinoline derivative 2a of this example is as follows:

2-chloro-3-benzimidazolylquinoline 1a (0.5mmol), ethanol (or toluene 2mL), and N, N-dimethylethylenediamine (0.15mL, 1.0mmol) were sequentially added to a 25mL round-bottomed flask with electromagnetic stirring, and the flask was placed in a heating mantle at 80 ℃ for reflux reaction for 12h (reaction monitored by TLC using Dichloromethane (DCM), Ethyl Acetate (EA), methanol (MeOH) as developing agents in a ratio V_DCM:V_EA:V_MeOH9:3:1), removing the solvent under reduced pressure, and separating and purifying by silica gel column chromatography (gradient elution, eluent: V)_DCM:V_MeOH100:1) to yield 100mg of compound 2a as a pale yellow solid in 60% yield.

The nuclear magnetic resonance data of the 2-substituted amino-3-benzimidazole substituted quinoline derivative 2a of this example are:

N¹-(3-(1H-benzo[d]imidazol-2-yl)quinolin-2-yl)-N²,N²-dimethylethane-1,2-diamine(2a):¹H NMR(400MHz,Chloroform-d)δ9.48(d,J＝5.8Hz,1H),7.94(s,1H),7.32–7.18(m,5H),7.02(dd,J＝6.1,3.2Hz,2H),6.93(ddd,J＝8.0,6.5,1.5Hz,1H),3.99(q,J＝5.4Hz,2H),2.96(t,J＝5.7Hz,2H),2.65(s,6H).¹³C NMR(100MHz,Chloroform-d)δ153.9,149.5,147.3,134.6,130.2,127.4,125.3,121.6,121.5,111.2,59.8,45.9,38.2。

example 2

A2-substituted amino-3-benzimidazole substituted quinoline derivative 2b has a structural formula as follows:

the preparation of 2-substituted amino-3-benzimidazole substituted quinoline derivative 2b of this example: the procedure was carried out in the same manner as in example 1 except for using 7-fluoro-2-chloro-3-benzimidazolylquinoline 1b in place of 2-chloro-3-benzimidazolylquinoline 1 a.

The NMR data for 2-substituted amino-3-benzimidazole substituted quinoline derivatives 2b of this example are:

N¹-(3-(1H-benzo[d]imidazol-2-yl)-7-fluoroquinolin-2-yl)-N²,N²-dimethylethane-1,2-diamine:¹H NMR(400MHz,Methanol-d4)δ8.54(s,1H),7.80–7.71(m,1H),7.65(dt,J＝7.0,3.5Hz,2H),7.33–7.27(m,3H),7.06(td,J＝8.7,2.5Hz,1H),3.95(t,J＝6.4Hz,2H),3.05(t,J＝6.4Hz,2H),2.64(s,6H).¹³C NMR(100MHz,Methanol-d4)δ166.7,155.8,150.4,149.9,149.8,136.3,131.1,130.9,123.7,120.1,112.7,112.5,112.0,112.0,112.0,109.8,59.3,45.0,38.9.

example 3

A2-substituted amino-3-benzimidazole substituted quinoline derivative 2c, which has a structural formula as follows:

the preparation of 2-substituted amino-3-benzimidazole substituted quinoline derivative 2c of this example: the procedure was carried out in the same manner as in example 1 except for using 2, 6-dichloro-3-benzimidazolylquinoline 1c in place of 2-chloro-3-benzimidazolylquinoline 1 a.

The 2c nmr data for the 2-substituted amino-3-benzimidazole substituted quinoline derivative of this example are:

N¹-(3-(1H-benzo[d]imidazol-2-yl)-6-chloroquinolin-2-yl)-N²,N²-dimethylethane-1,2-diamine:¹H NMR(400MHz,Chloroform-d)δ9.68(t,J＝5.7Hz,1H),7.84(s,1H),7.22(s,2H),7.19–7.15(m,2H),7.14(s,1H),7.10(dd,J＝8.9,2.2Hz,1H),7.02(s,2H),4.01(q,J＝5.5Hz,2H),3.05(t,J＝5.6Hz,2H),2.74(s,6H).¹³C NMR(100MHz,Chloroform-d)δ158.4,152.9,149.2,139.1,135.4,131.9,130.8,130.3,127.1,116.8,62.8,47.7,41.5,33.6.

example 4

A2-substituted amino-3-benzimidazole substituted quinoline derivative 2d has a structural formula as follows:

the preparation method of 2-substituted amino-3-benzimidazole substituted quinoline derivative 2d of this example: the same procedure as in example 1 was repeated except for using 7-bromo-2-chloro-3-benzimidazolylquinoline 1d in place of 2-chloro-3-benzimidazolylquinoline 1 a.

The 2d nmr data for the 2-substituted amino-3-benzimidazole substituted quinoline derivative of this example are:

N¹-(3-(1H-benzo[d]imidazol-2-yl)-7-bromoquinolin-2-yl)-N²,N²-dimethylethane-1,2-diamine:¹H NMR(400MHz,DMSO-d6)δ13.38(s,1H),10.01(t,J＝5.3Hz,1H),8.84(s,1H),7.77(d,J＝1.9Hz,1H),7.70(d,J＝8.5Hz,2H),7.39(dd,J＝8.4,2.0Hz,1H),7.34–7.27(m,2H),3.84(q,J＝6.0Hz,2H),2.92(t,J＝6.2Hz,2H),2.52(s,6H).¹³C NMR(100MHz,DMSO-d6)δ:154.8,149.8,148.9,135.8,130.4,127.8,125.4,124.5,121.0,112.0,57.8,44.9,38.3.

example 5

A2-substituted amino-3-benzimidazole substituted quinoline derivative 2e has a structural formula as follows:

the preparation of 2-substituted amino-3-benzimidazole substituted quinoline derivative 2e of this example: the same procedure as in example 1 was repeated except for using 6-methyl-2-chloro-3-benzimidazolylquinoline 1e in place of 2-chloro-3-benzimidazolylquinoline 1 a.

The 2-substituted amino-3-benzimidazole substituted quinoline derivative 2e nmr data of this example are:

N¹-(3-(1H-benzo[d]imidazol-2-yl)-6-methylquinolin-2-yl)-N²,N²-dimethylethane-1,2-diamine:¹H NMR(400MHz,DMSO-d6)δ13.51(s,1H),9.81(t,J＝5.5Hz,1H),8.86(s,1H),7.68(s,2H),7.56–7.51(m,2H),7.45(dd,J＝8.4,2.1Hz,1H),7.29(dq,J＝7.2,3.8Hz,2H),3.91(q,J＝6.0Hz,2H),3.10(t,J＝6.3Hz,2H),2.67(s,6H),2.44(s,3H).¹³C NMR(100MHz,DMSO-d6)δ153.9,150.3,146.3,135.8,133.2,131.6,127.4,125.8,122.3,111.5,57.7,44.3,37.7,21.2.

example 6

A2-substituted amino-3-benzimidazole substituted quinoline derivative 2f, which has a structural formula as follows:

the preparation of 2-substituted amino-3-benzimidazole substituted quinoline derivative 2f of this example: the same procedure as in example 1 was repeated except for using 6, 7-difluoro-2-chloro-3-benzimidazolylquinoline 1f in place of 2-chloro-3-benzimidazolylquinoline 1 a.

The 2f nmr data for the 2-substituted amino-3-benzimidazole substituted quinoline derivative of this example are:

N¹-(3-(1H-benzo[d]imidazol-2-yl)-6,7-difluoroquinolin-2-yl)-N²,N²-dimethylethane-1,2-diamine:¹H NMR(400MHz,DMSO-d6)δ13.28(s,1H),9.95(t,J＝5.1Hz,1H),8.70(s,1H),7.77–7.63(m,3H),7.49(dd,J＝12.4,7.6Hz,1H),7.30(dq,J＝7.3,4.0Hz,2H),3.72(q,J＝6.0Hz,2H),2.68(t,J＝6.3Hz,2H),2.35(s,6H).¹³CNMR(100MHz,DMSO-d6)δ154.5,149.8,147.6,145.7,145.6,145.1,135.01,118.3,118.7,114.4,114.3,112.3,112.2,111.7,111.7,58.3,45.6,39.1.

example 7

A 2-substituted amino-3-benzimidazole substituted quinoline derivative 3c, which has the structural formula:

the preparation of 2-substituted amino-3-benzimidazole substituted quinoline derivative 3c of this example: the same procedures used in the preparation of example 1 were repeated except that 2, 6-dichloro-3-benzimidazolylquinoline 1c was used in place of 2-chloro-3-benzimidazolylquinoline 1a, N, N-diethylpropylamine in place of 1- (2-aminoethyl) piperidine.

The nuclear magnetic resonance data of the 2-substituted amino-3-benzimidazole substituted quinoline derivative 3c of this example are:

3-(1H-benzo[d]imidazol-2-yl)-6-chloro-N-(2-(piperidin-1-yl)ethyl)quinolin-2-amine:¹H NMR(400MHz,DMSO-d6)δ13.28(s,1H),9.96(t,J＝5.0Hz,1H),8.70(s,1H),7.66(d,J＝77.9Hz,5H),7.30(d,J＝7.0Hz,2H),3.76(s,2H),2.77–2.53(m,4H),2.50(s,2H),1.67–1.42(m,6H).¹³C NMR(100MHz,DMSO-d6)δ154.5,149.8,146.8,135.0,131.2,127.9,126.92,126.0,122.8,112.5,57.2,54.2,25.9,24.2.

example 8

A2-substituted amino-3-benzimidazole substituted quinoline derivative 4a has a structural formula as follows:

the preparation method of 2-substituted amino-3-benzimidazole substituted quinoline derivative 4a of this example: using 2-chloro-3-benzimidazolylquinoline 1a as a starting material, N, N-diethylpropylamine was replaced with 1- (2-aminoethyl) piperidine, and the other steps were the same as in the preparation of example 1.

The NMR data for the 2-substituted amino-3-benzimidazole substituted quinoline derivative 4a of this example are:

3-(1H-benzo[d]imidazol-2-yl)-N-(2-(pyrrolidin-1-yl)ethyl)quinolin-2-amine

¹H NMR(400MHz,Chloroform-d)δ9.48(t,J＝5.5Hz,1H),7.91(s,1H),7.29–7.18(m,5H),7.04–6.98(m,2H),6.94–6.89(m,1H),3.99(q,J＝5.5Hz,2H),3.13(t,J＝5.7Hz,2H),3.00–2.93(m,4H),1.90(h,J＝3.0Hz,4H).¹³C NMR(100MHz,Chloroform-d)δ154.0,149.6,147.6,134.4,130.2,127.3,125.5,121.6,121.5,111.3,56.7,54.6,39.6,23.7.

example 9

A2-substituted amino-3-benzimidazole substituted quinoline derivative 4b has a structural formula as follows:

the preparation of 2-substituted amino-3-benzimidazole substituted quinoline derivative 4b of this example: the same procedures used in the preparation of example 1 were repeated except for using 7-fluoro-2-chloro-3-benzimidazolylquinoline 1b in place of 2-chloro-3-benzimidazolylquinoline 1a, and replacing N, N-diethylpropylamine with 1- (2-aminoethyl) piperidine.

The NMR data for the 2-substituted amino-3-benzimidazole substituted quinoline derivative 4b of this example are:

3-(1H-benzo[d]imidazol-2-yl)-N-(2-(pyrrolidin-1-yl)ethyl)quinolin-2-amine

example 10

A 2-substituted amino-3-benzimidazole substituted quinoline derivative 4c, which has a structural formula:

the preparation method of 2-substituted amino-3-benzimidazole substituted quinoline derivative 4c of this example: the same procedures as in example 1 were repeated except for using 2, 6-dichloro-3-benzimidazolylquinoline 1c in place of 2-chloro-3-benzimidazolylquinoline 1a, N, N-diethylpropylamine in place of 1- (2-aminoethyl) pyrrolidine.

The nuclear magnetic resonance data of the 2-substituted amino-3-benzimidazole substituted quinoline derivative 4c of this example are:

3-(1H-benzo[d]imidazol-2-yl)-6-chloro-N-(2-(pyrrolidin-1-yl)ethyl)quinolin-2-amine:¹H NMR(400MHz,Chloroform-d)δ7.94–7.85(m,1H),7.06(q,J＝6.5Hz,3H),6.88–6.63(m,5H),3.45(q,J＝6.5,6.1Hz,2H),2.87(dq,J＝20.6,6.9Hz,6H),1.57(s,4H).¹³C NMR(100MHz,Chloroform-d)δ158.0,152.8,149.3,138.9,135.1,131.6,130.7,130.3,127.1,127.0,127.0,127.0,127.0,116.4,59.5,58.3,42.6,27.2.

example 11

A2-substituted amino-3-benzimidazole substituted quinoline derivative 4d has a structural formula as follows:

the preparation method of 2-substituted amino-3-benzimidazole substituted quinoline derivative 4d of this example: the same procedures used in the preparation of example 1 were repeated except for using 7-bromo-2-chloro-3-benzimidazolylquinoline 1d in place of 2-chloro-3-benzimidazolylquinoline 1a, N, N-diethylpropylamine in place of 1- (2-aminoethyl) pyrrolidine.

The 4d nmr data for the 2-substituted amino-3-benzimidazole substituted quinoline derivative of this example are:

3-(1H-benzo[d]imidazol-2-yl)-7-bromo-N-(2-(pyrrolidin-1-yl)ethyl)quinolin-2-amine:¹H NMR(400MHz,DMSO-d6)δ13.58(s,1H),10.02(t,J＝5.8Hz,1H),8.96(s,1H),7.83(d,J＝1.9Hz,1H),7.74(dd,J＝18.0,8.3Hz,2H),7.65–7.58(m,1H),7.43(dd,J＝8.4,2.0Hz,1H),7.30(m,2H),4.05(q,J＝6.1Hz,2H),3.51(t,J＝6.5Hz,3H),1.95(s,4H).¹³C NMR(100MHz,DMSO-d6)δ154.8,149.8,148.6,142.8,136.1,134.4,130.5,128.0,125.8,124.6,124.2,122.7,121.3,119.3,112.1,111.9,54.0,53.6,37.5,23.2.

example 12

A2-substituted amino-3-benzimidazole substituted quinoline derivative 4e has a structural formula as follows:

the preparation of 2-substituted amino-3-benzimidazole substituted quinoline derivative 4e of this example: the same procedures used in the preparation of example 1 were repeated except for using 6-methyl-2-chloro-3-benzimidazolylquinoline 1e in place of 2-chloro-3-benzimidazolylquinoline 1a, N, N-diethylpropylamine in place of 1- (2-aminoethyl) pyrrolidine.

The nuclear magnetic resonance data of the 2-substituted amino-3-benzimidazole substituted quinoline derivative 4e of this example are:

3-(1H-benzo[d]imidazol-2-yl)-6-methyl-N-(2-(pyrrolidin-1-yl)ethyl)quinolin-2-amine:¹H NMR(400MHz,DMSO-d6)δ13.66(s,1H),9.84(t,J＝5.7Hz,1H),8.94(s,1H),7.76(d,J＝7.9Hz,1H),7.64(d,J＝7.9Hz,1H),7.55(d,J＝8.7Hz,2H),7.47(dd,J＝8.5,2.1Hz,1H),7.30(dt,J＝14.0,7.6Hz,2H),4.05(q,J＝6.2Hz,2H),3.52(t,J＝6.4Hz,2H),2.44(s,3H),2.00–1.92(m,4H).¹³C NMR(100MHz,DMSO-d6)δ153.8,150.2,146.1,142.8,136.0,134.5,133.3,131.9,127.5,125.9,123.9,122.5,122.4,119.2,111.9,111.6,54.0,53.9,37.6,23.2,21.2.

example 13

A2-substituted amino-3-benzimidazole substituted quinoline derivative 4f has a structural formula as follows:

the preparation of 2-substituted amino-3-benzimidazole substituted quinoline derivative 4f of this example: the same procedure as in example 1 was repeated except for using 6, 7-difluoro-2-chloro-3-benzimidazolylquinoline 1f in place of 2-chloro-3-benzimidazolylquinoline 1a, and replacing N, N-diethylpropylamine with 1- (2-aminoethyl) pyrrolidine.

The 4f nmr data for the 2-substituted amino-3-benzimidazole substituted quinoline derivative of this example are:

3-(1H-benzo[d]imidazol-2-yl)-6,7-difluoro-N-(2-(pyrrolidin-1-yl)ethyl)quinolin-2-amine:¹H NMR(400MHz,DMSO-d6)δ13.37(s,1H),9.99(t,J＝5.4Hz,1H),8.77(s,1H),7.77(dd,J＝10.8,9.0Hz,1H),7.67(h,J＝4.6Hz,2H),7.54(dd,J＝12.4,7.6Hz,1H),7.31(dt,J＝6.3,3.6Hz,2H),3.88(q,J＝6.0Hz,2H),3.21–2.95(m,6H),1.90–1.81(m,4H).¹³C NMR(100MHz,DMSO-d6)δ154.6,149.7,145.5,145.4,135.3,118.6,118.5,114.5,114.3,112.5,112.3,111.9,111.9,54.4,54.2,38.9,23.4.

example 14

4g of 2-substituted amino-3-benzimidazole substituted quinoline derivative, which has the structural formula:

the preparation of 4g of 2-substituted amino-3-benzimidazole substituted quinoline derivative of this example: the same procedures used in the preparation of example 1 were repeated except for using 1g of 6-bromo-2-chloro-3-benzimidazolylquinoline instead of 2-chloro-3-benzimidazolylquinoline 1a, N, N-diethylpropylamine instead of 1- (2-aminoethyl) pyrrolidine.

The 4g NMR data for the 2-substituted amino-3-benzimidazole substituted quinoline derivative of this example are:

3-(1H-benzo[d]imidazol-2-yl)-6-bromo-N-(2-(pyrrolidin-1-yl)ethyl)quinolin-2-amine¹H NMR(400MHz,DMSO-d6)δ13.78(s,1H),9.97(t,J＝5.8Hz,1H),9.01(s,1H),7.93(d,J＝2.3Hz,1H),7.80–7.71(m,2H),7.61(dd,J＝21.7,8.4Hz,2H),7.31(dt,J＝22.8,7.2Hz,2H),4.07(q,J＝6.3Hz,2H),3.52(t,J＝6.5Hz,3H),3.37(s,3H),1.96(s,4H).¹³C NMR(100MHz,DMSO-d6)δ154.4,149.7,146.5,142.7,135.6,134.5,133.9,130.1,128.3,124.2,123.9,122.7,119.3,114.6,112.6,112.0,53.9,53.6,37.5,23.2.

example 15

A2-substituted amino-3-benzimidazole substituted quinoline derivative 4h has a structural formula as follows:

the preparation of the 2-substituted amino-3-benzimidazole substituted quinoline derivative of this example was carried out for 4 h: the same procedures as in example 1 were repeated except for using 7-methoxy-2-chloro-3-benzimidazolylquinoline 1h instead of 2-chloro-3-benzimidazolylquinoline 1a, N, N-diethylpropylamine instead of 1- (2-aminoethyl) pyrrolidine.

The 4h nmr data for the 2-substituted amino-3-benzimidazole substituted quinoline derivative of this example are:

3-(1H-benzo[d]imidazol-2-yl)-7-methoxy-N-(2-(pyrrolidin-1-yl)ethyl)quinolin-2-amine:¹H NMR(400MHz,DMSO-d6)δ13.18(s,1H),9.92(t,J＝5.8Hz,1H),8.77(s,1H),7.72(dd,J＝16.0,8.3Hz,2H),7.61(d,J＝7.8Hz,1H),7.29(dq,J＝14.9,7.1Hz,2H),7.03–6.95(m,2H),4.04(q,J＝6.0Hz,2H),3.91(s,3H),3.59–3.32(m,6H),1.99(dt,J＝9.5,5.4Hz,4H).¹³C NMR(100MHz,DMSO-d6)δ162.3,154.9,150.4,149.6,142.8,136.0,134.4,123.0,123.8,122.5,119.1,117.3,115.0,111.7,109.0,105.5,55.9,54.7,54.4,37.7,23.1.

example 16

A2-substituted amino-3-benzimidazole substituted quinoline derivative 5 has a structural formula as follows:

the preparation of 2-substituted amino-3-benzimidazole substituted quinoline derivative 5 of this example: the same procedure as in example 1 was repeated except for using 2-chloro-3-dimethylbenzimidazolyl quinoline 1i in place of 2-chloro-3-benzimidazolylquinoline 1a, N, N-diethylpropylamine in place of 1- (2-aminoethyl) pyrrolidine.

The NMR data for 2-substituted amino-3-benzimidazole substituted quinoline derivative 5 of this example are:

3-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)-N-(2-(pyrrolidin-1-yl)ethyl)quinolin-2-amine:¹H NMR(400MHz,DMSO-d6)δ13.50(s,1H),9.99(t,J＝5.7Hz,1H),9.02(s,1H),7.76(d,J＝8.0Hz,1H),7.62(q,J＝4.6,3.1Hz,2H),7.53(s,1H),7.39(s,1H),7.30(ddd,J＝8.1,5.4,2.6Hz,1H),4.06(q,J＝6.2Hz,2H),3.51(t,J＝6.5Hz,2H),2.35(d,J＝5.6Hz,6H),2.01–1.92(m,4H).¹³C NMR(100MHz,DMSO-d6)δ154.1,149.2,147.6,141.5,135.9,133.0,132.8,131.0,128.5,126.0,122.8,122.6,119.2,111.99,53.8,53.7,37.5,23.2,20.5.

example 17

A2-substituted amino-3-benzimidazole substituted quinoline derivative 6 has a structural formula as follows:

the preparation of 2-substituted amino-3-benzimidazole substituted quinoline derivative 6 of this example: the same procedures as in example 1 were repeated except for using 2-chloro-3-dimethylbenzimidazolyl quinoline 1i in place of 2-chloro-3-benzimidazolylquinoline 1a, N, N-diethylpropylamine in place of 1- (2-aminoethyl) piperidine.

The NMR data for the 2-substituted amino-3-benzimidazole substituted quinoline derivative 6 of this example are:

3-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)-N-(2-(piperidin-1-yl)ethyl)quinolin-2-amine:¹H NMR(400MHz,DMSO-d6)δ13.33(s,1H),9.98(s,1H),8.94(s,1H),7.77(d,J＝8.0Hz,1H),7.63(d,J＝5.3Hz,2H),7.46(d,J＝49.2Hz,2H),7.30(t,J＝6.1Hz,1H),4.10(q,J＝6.3Hz,2H),3.42(d,J＝13.1Hz,6H),2.36(s,6H),1.82(q,J＝6.6,6.1Hz,4H),1.56(s,2H).¹³C NMR(100MHz,DMSO-d6)δ154.2,149.1,135.9,131.1,128.6,125.9,122.9,122.6,111.9,55.9,53.0,36.1,22.9,21.8,20.5.

example 18

A2-substituted amino-3-benzimidazole substituted quinoline derivative 7 has a structural formula as follows:

the preparation of 2-substituted amino-3-benzimidazole substituted quinoline derivative 7 of this example: the procedure was carried out in the same manner as in example 1 except for using 2-chloro-3-dimethylbenzoimidazolyl quinoline 1i in place of 2-chloro-3-benzimidazolyl quinoline 1 a.

The NMR data for the 2-substituted amino-3-benzimidazole substituted quinoline derivative 7 of this example are:

N¹-(3-(5,6-dimethyl-1H-benzo[d]imidazol-2-yl)quinolin-2-yl)-N²,N²-dimethylethane-1,2-diamine:¹H NMR(400MHz,DMSO-d6)δ13.33(s,1H),10.00(s,1H),8.95(s,1H),7.77(d,J＝8.0Hz,1H),7.69–7.58(m,2H),7.56–7.35(m,2H),7.31(ddd,J＝8.1,6.0,2.1Hz,1H),4.07(q,J＝5.8Hz,2H),3.47(t,J＝6.2Hz,2H),2.94(s,6H),2.36(s,6H).¹³C NMR(100MHz,DMSO-d6)δ154.3,149.1,147.4,135.9,131.2,128.6,125.9,122.9,122.6,111.9,56.9,43.1,36.6,20.5.

application of the Compound of example 1 to topoisomerase I inhibitory Activity and DNA topoisomerase I mediated relaxation experiments

2-substituted amino-3-benzimidazole substituted quinoline derivatives (hereinafter referred to as compounds) 2a, 2b, 2c, 2d, 2f, 3c, 4a, 4b, 4c, 4d, 4e, 4f, 4g, 4h, 5, 6,7 and the like were subjected to an inhibition experiment on DNA topoisomerase I activity by agarose gel electrophoresis.

The experimental method comprises the following specific steps:

(1) preparing 1% agarose gel solution: weighing a certain amount of agarose, placing the agarose in a triangular flask, adding 1xTAE buffer solution, placing the agarose in a microwave oven, heating the agarose until the agarose is completely dissolved and is transparent, taking out the agarose and shaking the agarose to ensure that the agarose is completely dissolved and no solid particles exist;

(2) preparation of agarose gel: cooling the gel to about 60 ℃, slightly shaking, uniformly mixing, pouring the agarose gel into a preparation plate, inserting a comb, cooling at room temperature for 30-60 min until the gel is completely solidified, carefully pulling out the comb, placing the gel in an electrophoresis tank, and adding a proper amount of electrophoresis buffer solution to ensure that the gel is about 2-3 mm higher than the gel;

(3) sample incubation and electrophoresis: taking 250ng of pBR322 plasmid DNA, a compound to be detected with different concentrations, 1 muL of 0.1% BSA, 1 muL of 10xTopoI buffer solution and Topo I, adding deionized water to the final volume of 10 muL, incubating in a water bath at 37 ℃ for 30min, then incubating in a water bath at 65 ℃ for 5min to terminate the reaction, after the reaction is terminated, respectively adding 2 muL of 6xDNA loading buffer solution into a sample, taking the pBR322 plasmid DNA alone as a negative control, taking camptothecin as a positive control, finally loading the sample into a gel, and carrying out electrophoresis for about 1.5 h;

(4) and (3) developing and imaging: after electrophoresis, the gel block was incubated in 1mg/L Ethidium Bromide (EB) for 30min and then imaged on a gel system.

The experimental results are as follows: FIG. 3 shows the results of Topo I inhibitory activity of the compounds at 100. mu.M, and FIG. 4 shows the results of experiments on the Topo I inhibitory pattern of the compounds. As can be seen from FIGS. 3 and 4, most of the compounds exhibited strong Topo I inhibitory activity, and their mechanisms of action were different from those of Topo I poisoning agent, camptothecin, and Topo I enzyme inhibitor, luteolin. As can be seen in fig. 4(B, E), both the topoisomerase poison camptothecin and the topoisomerase catalytic inhibitor luteolin, which demonstrate the ability of the test compound to interfere with the enzymatic activity of the enzyme, are strongly inhibitory, but it is not clear whether the selected compound is a poison or an enzyme inhibitor; as shown in fig. 4(C, F), camptothecin is a topoisomerase poison, which acts by forming a stable trimodal complex with DNA and compound, but there is no such condition and thus no phenomenon of inhibiting topoisomerase, and the reported inhibitory factor luteolin acts directly on topoisomerase to inhibit its activity in the absence of DNA in the presence of catalase, so our compound produces the same phenomenon as luteolin, which primarily indicates that compounds 2a,4C are inhibitors and not poisons; as shown in FIG. 4(D, G), none of camptothecin and luteolin showed inhibitory activity, while selected compounds showed superior inhibitory activity, and it is hypothesized that the compounds had DNA intercalation effects that would cause the uncoiled DNA to become a tight negative supercoil again allowing it to re-swim, which is consistent with the results of the subsequent supercoiled DNA intercalation experiment. Therefore, it can be said that our compounds have dual actions of topoisomerase inhibitors and DNA intercalation, and together have strong topoisomerase inhibitory effects.

Application example 2 cleavable Complex experiment

The experimental method comprises the following steps: supercoiled DNA (pBR322, 100 ng; Thermo, USA) was reacted with 3 units of topo I (TopoGen, USA) for 10 minutes, followed by addition of the test compound. The reaction mixture was incubated in a 37 ℃ water bath for 20 minutes; the reaction was stopped with 0.5% sarcosyl and then digested with proteinase K at 45 ℃ for 30min to remove excess protein; after addition of the loading buffer, the reaction mixture was heated in a water bath at 70 ℃ for 2 minutes, and then subjected to electrophoresis on a 1.0% agarose gel in TAE buffer containing 0.5ug/mL ethidium bromide, and then the gel was decolorized with water for 20 minutes, and DNA bands were visualized by UV transillumination.

The experimental results are as follows: electrophoresis is carried out by using agarose gel containing ethidium bromide, if DNA is cut off, a marked band is shown as a Nick band, FIG. 3 is an experimental electrophoresis chart of Topo 1-DNA cleavable complex of compound 2a, and it can be seen from the graph that compared with CPT, the insertion effect of EB is gradually saturated by DNA moving to the anode, and the insertion of EB can make the untwisted DNA structure compact and make the migration speed faster; otherwise, the electrophoretic mobility of the corresponding band of nicked DNA due to the stability of the cleavable complex is not affected by the presence of ethidium bromide but stays at the origin, and the results obtained by incubating DNA and enzyme with compound 2a at concentrations from 5 μ M to 50 μ M) indicate that this compound is not able to induce the formation of a cleavable complex with a lytic agent. For the poisoning agent CPT, a large amount of nicked DNA had formed at a concentration of 0.5. mu.M, further demonstrating that this compound is not a topoisomerase poisoning agent.

Application example 3 comet experiment

The experimental method comprises the following steps: comet assays were performed using single cell gel electrophoresis and Trevigen kit (gaithersburg, usa). T24 cells were seeded at a density of 1 × 105 cells per well in a six-well plate, treated with 10 μ M CPT and compound 2a for 24 hours, and then the collected cells were resuspended in ice-cold PBS. 1% Normal Melting Agarose (NMA) was melted in a water bath, spread on a glass slide, and cured at 4 ℃ for 10 minutes in the dark. Then 10. mu.L of the resuspended cells were mixed with 80. mu.L of 0.7% Low Melting Agarose (LMA), spread on a slide, and solidified at 4 ℃ in the dark for 10 minutes; after the second layer of low melting point gel is solidified, 75 mu L of low melting point agarose is carefully added, and the mixture is solidified for 10min in a dark place at 4 ℃; the lysates were lysed at 4 ℃ in ice-cold conditions for 2h in the dark, run at 15V for 20 min in alkaline conditions, the slides were rinsed twice with distilled water and then neutralized with 0.4mm tris-HCl (PH 7.5) for 10min, PI stained at 4 ℃ in the dark for 5min, and comet images were obtained using an inverted fluorescence microscope (Zeiss, Axiovert 200) at 20 x magnification.

The experimental results are as follows: whether compound 2a induces DNA damage like a topo-toxicant was assessed by the extent of comet (tail) formation indicating the extent of DNA strand breaks. Fig. 4 shows the comet images obtained by the test, 2a does not generate obvious tail representing damage, only cell shrinkage occurs after dosing compared with a blank group, and the well-known Topo I poison CPT remarkably induces DNA damage to generate obvious comet (tail), and the result shows that the tested compound does not cause damage to DNA and has a different action mechanism from a poisoning agent.

Application example 4 in vitro antiproliferative Activity test experiment

The experimental method comprises the following steps:

(1) culturing test cells HepG2 (liver cancer cells), T-24 (bladder cancer cells), HELA (cervical cancer cells), NCI-H460 (lung cancer cells) and HL-7702 (human normal liver cells) to logarithmic phase, digesting with trypsin, blowing the cells evenly with a culture solution containing 10% fetal calf serum to prepare a single cell suspension, adding a proper culture medium, uniformly mixing, inoculating into a 96-well plate, wherein each well volume is 180 mu L, and 200 mu L of PBS is added around the 96-well plate to reduce the evaporation of the drug and the culture medium;

(2) placing 96-well plate at 37 deg.C and 5% CO₂Incubating in an incubator until a cell monolayer is attached to the bottom of a well and grows to 70-80%, adding a compound with a concentration gradient (1, 2, 4, 6, 8 mu M) (10 mM is prepared by dissolving the compound in DMSO, and diluting the compound with PBS to the required concentration); adding 20 μ L of the solution into each well, setting 5 multiple wells at each concentration to reflect the real condition, and setting control wells (cells, culture solution, MTT, dimethyl sulfoxide)Sulfone, no additional compound);

(3) placing the mixture in a cell culture box for continuous culture for 48 hours, adding 10 mu L of MTT solution (5mg/mL) into each hole, and culturing in the culture box for 4-6 hours;

(4) terminating the culture, carefully removing culture solution in the wells, adding 100 μ L of dimethyl sulfoxide (DMSO) into each well, and placing in a micro-oscillator for low-speed oscillation for 10min to fully dissolve the crystals; measuring absorbance values of all the holes in an enzyme-linked immunosorbent assay detector, and setting a test wavelength to be 570nm and a reference wavelength to be 630 nm;

(5) inhibition rate and IC₅₀Calculation of the value:

inhibition/% (1-mean OD value of sample/mean OD value of control) × 100%, IC₅₀The values were obtained by statistical analysis of the SPSS software; the experimental results were obtained from 3 independent replicates, and the data are presented as mean ± standard deviation.

The experimental results are as follows: table 1 shows the IC of the compounds on different cell lines₅₀As can be seen from the table, the series of compounds all show good proliferation inhibition activity on human gastric cancer cells MGC-803, lung cancer cells NCI-H460, bladder cancer cells T-24 and cervical cancer cells HeLa, except compounds 4d and 4H, the other compounds all show good inhibition activity on human liver cancer cells HepG2, and the toxicity of all the compounds on human normal liver cells HL-7702 is obviously less than that of a clinical antitumor drug Adriamycin (ADM).

TABLE 1 IC of Compounds on different cell lines₅₀(μM)

In vivo antitumor Activity test experiment of Compound 2a of application example 5

The experimental method comprises the following steps:

(1) constructing an animal model:

t-24 tumor cells in the logarithmic growth phase were taken and inoculated into 30 nude mice (SPF-grade BALB/c nude mice, supplied by Kyowa Kavens laboratory animals Co., Ltd. (laboratory animal production license: SCXK (threo) 2016-:SCXK (su) 2017-. The age in days: 4-6 weeks at the time of purchase, 6-8 weeks at the time of administration; weight: the weight is 16-18g when purchasing and 18-20g when starting administration; sex: male; number of animals per group: each group had 6. ) Subcutaneous right axillary fossa, cell inoculation amount of 5X 10⁶Measuring the diameter of the transplanted tumor with vernier caliper until the tumor grows to 100mm³On the other hand, 24 tumor-bearing nude mice with good growth status and good uniformity of tumor size were selected and randomly divided into 4 groups of 6 mice each, namely, a model group, a 2a low dose group (10mg/kg), a 2a high dose group (20mg/kg), and a positive drug doxorubicin group (3 mg/kg).

(2) The preparation and administration method of the medicine comprises the following steps:

2a Low dose group (10 mg/kg): weighing 9.6mg of 2a powder, dissolving the powder in 10 mu L of DMSO, completely dissolving the powder, and adding physiological saline to ensure that the total volume reaches 9.6mL and the concentration is 1 mg/mL; the medicine is administrated by intraperitoneal injection for 1 time every other day, the experimental time is 16 days, the medicine is administrated for 8 times totally, and the administration volume is 0.2mL/20g body weight;

2a high dose group (20 mg/kg): weighing 19.2mg of 2a powder, dissolving the powder in 20 mu L of DMSO, completely dissolving the powder, and adding physiological saline to ensure that the total volume reaches 9.6mL and the concentration is 2 mg/mL; the medicine is administrated by intraperitoneal injection for 1 time every other day, the experimental time is 16 days, the medicine is administrated for 8 times totally, and the administration volume is 0.2mL/20g body weight;

positive drug doxorubicin group (3 mg/kg): weighing 3.2mg of adriamycin powder, dissolving the adriamycin powder in 10 mu L of DMSO, completely dissolving the adriamycin powder, and adding physiological saline to ensure that the total volume reaches 9.6mL and the concentration is 1 mg/mL; the medicine is administrated by intraperitoneal injection for 1 time every other day, the experimental time is 16 days, the medicine is administrated for 8 times totally, and the administration volume is 0.2mL/20g body weight;

(3) processing and analyzing experimental results:

dynamically observing the anti-tumor effect of the tested object by using a method for measuring the tumor diameter, wherein the tumor diameter is measured once every other day, and the weight of the nude mouse is weighed while the tumor diameter is measured; the mice were sacrificed by removing the neck 16 days after administration, tumor masses were surgically removed and weighed, and tumor tissues were fixed with 10% neutral formalin;

organ coefficient (%) (organ wet weight/body weight) × 100;

the formula for Tumor Volume (TV) is:

TV＝1/2×a×b²

wherein a and b respectively represent length and width;

calculating Relative Tumor Volume (RTV) according to the measurement result, wherein the calculation formula is as follows: RTV ═ V_t/V₀(ii) a Wherein V₀When administered separately from the cage (i.e. d)₀) Measurement of the resulting tumor volume, V_tFor tumor volume at each measurement; the evaluation index of the antitumor activity is relative tumor proliferation rate T/C (%), and the calculation formula is as follows:

T/C(％)＝TRTV/CRTV×100

TRTV: treatment group RTV; CRTV: negative control group RTV.

The experimental results are as follows: tables 2 and 5 show the tumor weight and tumor inhibition rate of each group of tumor-bearing nude mice, tables 3 and 8 show the effect of the change in the body weight of each group of tumor-bearing nude mice, fig. 6 shows a photograph of each group of tumor-bearing nude mice and a photograph of each group of tumors, and fig. 7 shows a photograph of HE-stained organ sections of each group of tumor-bearing nude mice. From the above results, it is clear that the tumor volume of the nude mice xenografted after the compound 2a was exposed to the model group was significantly reduced, and the tumor volume reduction of the high dose group was larger than that of the low dose group, showing a certain dose-dependent relationship. The activity intensity of the high-dose group (20mg/kg) of the compound 2a for inhibiting the tumor growth is equivalent to that of the positive medicine group (5 mg/kg). Although the dose was higher than the group of positive drugs, the tested compounds produced less toxicity to mice than the positive drug doxorubicin. The compound 2a is administrated once every other day in a low-dose group of 10mg/kg and a high-dose group of 20mg/kg in an intraperitoneal injection way for 8 times, and results show that the tumor inhibition rates of the compound 2a on the transplanted tumor of a gastric cancer T-24 nude mouse of a human body are respectively 40.0 percent and 54.3 percent, the cancer inhibition effect is obvious, and no obvious organ injury is generated on the mouse.

TABLE 2 tumor weight and tumor inhibition ratio of tumor-bearing nude mice (X + -SD, n ═ 6, body weight: g) of each group

Group of	Dosage (mg/kg)	Number of initial animals	Number of terminal animals	Tumor weight (g)	Tumor inhibition Rate (%)
						Model set		6	6	1.633±0.150	-
2a Low dose group	10	6	6	0.980±0.119^****	40
						2a high dose group	20	6	6	0.746±0.075^****	54.3
Positive drug group	5	6	6	0.626±0.084^****	61.7

Note: in comparison with the set of models,^****p<0.0001。

TABLE 3 influence of weight change in groups of tumor-bearing nude mice (X + -SD, n ═ 6, weight: g)

Group of

Dosage (mg/kg)

1d

3d

5d

7d

9d

Model set

—

19.6±0.6

19.8±0.5

19.9±0.5

20.1±0.6

20.3±0.6

2a Low dose group

10

19.5±0.8

19.7±0.7

19.8±0.7

20.0±0.7

20.1±0.8

2a high dose group

20

19.3±0.5

19.5±0.5

19.6±0.4

19.8±0.4

20.0±0.4

Positive drug group

5

19.5±0.9

19.6±0.9

19.8±0.9

19.9±0.9

20.0±0.9

Group of

Dosage (mg/kg)

11d

13d

15d

Model set

—

20.4±0.6

20.6±0.6

20.8±0.6

2a Low dose group

10

20.3±0.8

20.5±0.8

20.6±0.8

2a high dose group

20

20.2±0.5

20.4±0.5

20.5±0.4

Positive drug group

5

20.1±0.9

20.2±0.8

20.3±0.8

In conclusion, the 2-substituted amino-3-benzimidazole substituted quinoline derivative has the characteristics of antitumor activity, high efficiency and low toxicity as a strong topoisomerase I inhibitor, and can be used for developing novel topoisomerase I inhibitors and lead compounds of antitumor drugs.

The foregoing descriptions of specific exemplary embodiments of the present invention have been presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and its practical application to enable one skilled in the art to make and use various exemplary embodiments of the invention and various alternatives and modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.

Claims

1. A2-substituted amino-3-benzimidazole substituted quinoline derivative is characterized in that the structural formula is as follows:

2. The process for producing a 2-substituted amino-3-benzimidazole substituted quinoline derivative according to claim 1, wherein the compound of formula (II) is dissolved in an organic solvent, and then the compound of formula (III) is added to the solution and heated to reflux; after the reaction is finished, decompressing to remove the solvent, separating and purifying to obtain the target compound shown in the formula (I), namely the 2-substituted amino-3-benzimidazole substituted quinoline derivative;

in the formula, R¹Is hydrogen, fluorine, chlorine, bromine, methyl or methoxy, R is dimethylamino, piperidine or tetrahydropyrrole, R is²Is hydrogen or methyl, and n is 2.

3. The process according to claim 2, wherein the compound of formula (II): the molar ratio of the compound of the formula (III) is 1: 1.5-2.0.

4. The method according to claim 2, wherein the organic solvent is ethanol or toluene.

5. The preparation method according to claim 2, wherein the reaction temperature is reflux temperature, and the reaction time is 10-12 h.

6. The method according to claim 2, wherein the heating reaction is carried out under electromagnetic stirring conditions.

7. The preparation method according to claim 2, wherein the separation and purification is silica gel column chromatography separation and purification, gradient elution is performed, the eluent is dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is 100: 1.

8. Use of a 2-substituted amino-3-benzimidazole substituted quinoline derivative according to claim 1 for the development of novel topoisomerase I inhibitors.

9. Use of a 2-substituted amino-3-benzimidazole substituted quinoline derivative according to claim 1 for the preparation of topoisomerase I based antitumor drugs.