阅读说明：本技术 一种n-乙酰诺氟沙星的丙烯酮衍生物及其制备方法和应用 (Acrylone derivative of N-acetylnorfloxacin, and preparation method and application thereof ) 是由 武毅君 胡国强 于 2019-11-20 设计创作，主要内容包括：本发明公开了一种N-乙酰诺氟沙星的丙烯酮衍生物及其制备方法和应用,采用如下式Ⅰ化学结构通式：式I中,芳香环Ar为苯环或取代苯环或呋喃环或吡啶环。本发明的一种N-乙酰诺氟沙星的丙烯酮衍生物,实现了氟喹诺酮骨架与丙烯酮骨架的有效拼合,进而构筑了新的氟喹诺酮“类查尔酮”化合物,从而增加了新化合物的抗肿瘤活性及抗耐药性,并降低对正常细胞的毒副作用,可以作为抗肿瘤活性物质开发全新结构的抗肿瘤药物。(The invention discloses an acrylketone derivative of N-acetylnorfloxacin, a preparation method and application thereof, wherein the chemical structural general formula of the derivative is as follows:)

1. The propenone derivative of the N-acetylnorfloxacin is characterized by being a typical compound with the following structure:

2. the process for preparing the propenone derivative of N-acetylnorfloxacin according to claim 1, which comprises the following steps:

1) taking N-acetylnorfloxacin shown in a formula II as a raw material, reacting with Carbonyldiimidazole (CDI) to prepare an N-acetylnorfloxacin imidazole amide compound shown in a formula III, and then carrying out condensation reaction with monoethyl malonate potassium salt to prepare a C-3 formyl ethyl acetate compound of the N-acetylnorfloxacin shown in a formula IV; finally, the N-acetyl norfloxacin C-3 ethanone shown in the formula V is prepared by hydrolysis and decarboxylation reaction of the formula IV:

2) the propenone derivative of N-acetyl norfloxacin as shown in claim 1 can be prepared by carrying out Claisen-Schmidt condensation reaction on N-acetyl norfloxacin C-3 ethanone shown in a formula V and aromatic aldehyde under the catalysis of alkali to form an propenone structure and carrying out post treatment.

3. The method for preparing the acrylketone derivative of N-acetylnorfloxacin according to claim 2, wherein the molar ratio of N-acetylnorfloxacin represented by the formula II to CDI is 1: 1.0-2.0, the molar ratio of N-acetylnorfloxacin imidazole amide represented by the formula III to monoethyl malonate potassium salt is 1: 1.0-1.5, and the molar ratio of N-acetylnorfloxacin-3 ethanone represented by the formula V to aromatic aldehyde is 1: 1.0-2.0.

4. The use of the acrylketone derivative of N-acetylnorfloxacin as defined in claim 1 for the preparation of an antitumor medicament.

5. The use of the acrylketone derivative of N-acetylnorfloxacin as claimed in claim 4, wherein the antitumor drug is a drug for treating non-small cell lung cancer, kidney cancer, liver cancer, stomach cancer, pancreatic cancer or leukemia.

Technical Field

The invention belongs to the technical field of innovative drug synthesis, and particularly relates to an acrylketone derivative of N-acetylnorfloxacin, a preparation method of the acrylketone derivative of N-acetylnorfloxacin, and application of the acrylketone derivative in antitumor drugs.

Background

New drug innovation stems from the discovery of leads, and rational drug molecular design based on structure or mechanism is an effective method for discovering leads. In the drug effect groups with various structures, the acrylketone structure is not only the characteristic structure of a chalcone compound which is a natural effective component, but also the characteristic drug effect group of a targeted antitumor drug sunitinib. Therefore, compounds constructed with acrylketone as a structural fragment and having various pharmacological activities have been attracting attention. However, most of natural chalcone compounds are multi-hydroxyl benzene ring substituted propenone compounds, and the poor water solubility of the compounds causes low bioavailability and limits clinical application. In addition, the topoisomerase which is an action target point of the antibacterial fluoroquinolone medicine is also an important action target point of the antitumor medicine, the antibacterial activity of the antibacterial fluoroquinolone medicine can be converted into the antitumor activity, and the fluoroquinolone C-3 carboxyl is not a pharmacophore required by the antitumor activity and can be replaced by a biological electron isostere to improve the antitumor activity of the fluoroquinolone medicine. However, the research on the replacement of the C-3 carboxyl group of fluoroquinolone with aryl acrylketone has not been reported. Based on the above, in order to improve the water solubility of chalcone, hydrophilic piperazinyl is introduced to increase the water solubility and improve the bioavailability and the activity of the chalcone, the invention uses the skeleton of the dominant pharmacophore of the fluoroquinolone drug N-acetylnorfloxacin, namely '1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one', as a substituent of an aryl propenone structure, and further designs the fluoroquinolone 'chalcone-like' derivative with a novel structure.

Therefore, the invention aims to provide the acrylketone derivative of the N-acetylnorfloxacin, which has the anti-tumor effect and efficacy and also provides a preparation method of the acrylketone derivative of the N-acetylnorfloxacin.

In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: the acrylketone derivative of N-acetylnorfloxacin has a chemical structural formula shown as a general formula I:

in the formula I, Ar is a benzene ring or a substituted benzene ring or a furan ring or a pyridine ring, and the compound is a compound with the following specific structure:

the preparation method of the acrylketone derivative of N-acetylnorfloxacin is prepared by using commercially obtained N-acetylnorfloxacin shown as a formula II as a raw material;

the preparation method comprises the following specific steps:

1) the N-acetyl norfloxacin imidazole amide compound shown in the formula III is prepared by taking N-acetyl norfloxacin shown in the formula II as a raw material and reacting with Carbonyldiimidazole (CDI), and the specific preparation method is as follows:

20g (55.4mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-carboxylic acid II is dissolved in 500mL of anhydrous acetonitrile, 15.2g (94.0mmol) of carbonyldiimidazole is added, and the reaction mixture is stirred in a water bath and refluxed until the starting material II disappears. Standing at room temperature, filtering to collect the generated solid, and recrystallizing with acetone to obtain a light yellow crystal of formula III, wherein the yield is 87.6%, and m.p.236-238 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.57(3H，t，CH₃)，2.18(3H，s，COCH₃) 2.67 to 3.46(8H, m, piperazine-H), 4.42(2H, q, N-CH)₂) 6.73-7.85 (3H, m, imidazole-H and 8-H), 8.03(1H, d, 5-H), 8.26(1H, s, imidazole-H), 8.85(1H, s, 2-H); MS (m/z): 412[ M + H]⁺Calculating (C)₂₁H₂₂FN₅O₃)：411.44。

As a further improvement, the molar ratio of the N-acetylnorfloxacin shown in the formula II to the carbonyldiimidazole is 1: 1.0-2.0, and the solvent can be at least one of acetonitrile, tetrahydrofuran, dioxane and dimethylformamide or a mixed solvent of the two.

2) The N-acetyl norfloxacin imidazole amide shown in a formula III and monoethyl malonate potassium salt are subjected to condensation reaction under the catalysis of triethylamine-magnesium chloride to prepare the C-3 formyl ethyl acetate compound of the N-acetyl norfloxacin shown in a formula IV, and the specific preparation method is as follows:

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- (1H-imidazol-1-yl) -quinolin-4 (1H) -one of formula III 16g (39.0mmol), magnesium chloride 6.6g (69.1mmol) and potassium monoethyl malonate 8.3g (49.0mmol) were added successively to 600mL of anhydrous acetonitrile, and 12.2g (12.0mmol) of triethylamine was added dropwise with stirring in an ice bath, and the reaction mixture was stirred in a water bath and refluxed until the starting material III disappeared. And (3) evaporating the solvent under reduced pressure, adding 500mL of water, adjusting the pH value to be approximately equal to 5.0 by using glacial acetic acid, filtering and collecting the solid, washing the solid to be neutral by using water, and recrystallizing the solid by using absolute ethyl alcohol to obtain an off-white crystal shown as a formula IV, wherein the yield is 62.4 percent, and m.p.227-229 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.32，1.58(6H，2t，2×CH₃)，2.14(3H，s，COCH₃) 2.76 to 3.56(8H, m, piperazine-H), 4.20(2H, s, COCH)₂CO)，4.26(2H，q，CO₂CH₂)，4.48(2H，q，N-CH₂)，6.74(1H，d，8-H)，8.05(1H，d，5-H)，8.87(1H，s,2-H)；MS(m/z)：432[M+H]⁺Calculating (C)₂₂H₂₆FN₃O₅)：431.47。

3) The C-3 ethyl formylacetate compound of the N-acetylnorfloxacin shown in the formula IV is hydrolyzed and decarboxylated by using 2 percent of sodium hydroxide aqueous solution by mass fraction, so that the C-3 ethanone compound of the N-acetylnorfloxacin shown in the formula V can be conveniently prepared, and the specific preparation method is as follows:

taking 10g (23.2mmol) of ethyl 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) quinolin-4 (1H) -one-3-formylacetate, suspending in 200mL of sodium hydroxide aqueous solution with the mass fraction of 2%, stirring in an oil bath and refluxing to react until the raw material IV disappears. Standing at room temperature, filtering to collect the generated solid, washing with water to be neutral, drying, and recrystallizing with absolute ethyl alcohol to obtain a light yellow crystal of formula V, wherein the yield is 75.3%, and m.p.234-236 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.56(3H，t，CH₃)，2.18(3H，s，N-COCH₃)，2.38(3H，s，COCH₃) 2.66 to 3.57(8H, m, piperazine-H), 4.48(2H, q, N-CH)₂)，6.76(1H，d，8-H)，8.03(1H，d，5-H)，8.86(1H，s,2-H)；MS(m/z)：360[M+H]⁺Calculating (C)₁₉H₂₂FN₃O₃)：359.40。

4) C-3 ethanone of N-acetylnorfloxacin shown in a formula V and aromatic aldehyde are subjected to Claisen-Schmidt aldol condensation reaction in absolute ethyl alcohol under the catalysis of alkali, and after the reaction is completed, a target compound is obtained by treatment, wherein the target compound is shown in a formula I, and the specific process is as follows:

wherein Ar in the formula I is a benzene ring or a substituted benzene ring or a furan ring or a pyridine ring.

The general synthetic preparation procedure for the target compound of formula i is: 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and the aromatic aldehyde (3.0mmol) and the base catalyst piperidine (0.1mL) were added. And (3) refluxing and reacting the mixed reactants for 15-24 h, standing at room temperature, filtering and collecting the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a light yellow crystal shown in the formula I.

As a further improvement, the mol ratio of the N-acetyl norfloxacin C-3 ethanone shown in the formula V to the aromatic aldehyde is 1: 1.0-1.5.

The base catalyst is at least one of piperidine, pyridine, triethylamine, morpholine, potassium acetate, sodium hydroxide ethanol solution or potassium hydroxide ethanol solution.

The application of the acrylketone derivative of N-acetylnorfloxacin in preparing antitumor drugs.

The anti-tumor drug is a drug for treating human non-small cell lung cancer, kidney cancer, liver cancer, stomach cancer, pancreatic cancer or leukemia.

The acrylketone derivative of N-acetylnorfloxacin disclosed by the invention is designed and synthesized by effectively combining a fluoroquinolone skeleton and an aryl acrylketone pharmacophore based on the split principle of the pharmacophores, and the complementation and activity superposition of the pharmacophores with different structures are realized, so that the effects of synergism, toxicity reduction and drug resistance are achieved, and the acrylketone derivative of N-acetylnorfloxacin can be developed as an anti-tumor drug with a brand new structure.

Detailed Description

Example 1

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3-cinnamoyl-quinolin-4 (1H) -one (I-1) having the chemical formula:

namely, Ar in the formula I is phenyl.

The preparation method of the compound comprises the following steps: 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -1-yl-quinolin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.40g (3.8mmol) of benzaldehyde and piperidine, a base catalyst (0.1mL) were added. And (3) refluxing and reacting the mixed reactants for 18h, standing at room temperature, filtering and collecting the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a light yellow crystal, namely a formula I-1, wherein the yield is 86.2%, and m.p.235-237 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.58(3H，t，CH₃)，2.23(3H，s，COCH₃) 2.76 to 3.57(8H, m, piperazine-H), 4.36(2H, q, N-CH)₂) 6.75 to 7.82(7H, m, 8-H, Ph-H and 2' -H),8.26(1H，d，5-H)，8.53(1H，d，3′-H),8.86(1H，s，2-H)；MS(m/z)：448[M+H]⁺calculating (C)₂₆H₂₆FN₃O₃)：447.51。

Example 2

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- (4-methoxycinnamoyl) -quinolin-4 (1H) -one (I-2) having the chemical formula:

namely, Ar in the formula I is p-methoxyphenyl.

The preparation method of the compound comprises the following steps: 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.57g (4.2mmol) of 4-methoxybenzaldehyde and piperidine, a basic catalyst (0.1mL) were added. And (3) carrying out reflux reaction on the mixed reactants for 20h, standing at room temperature, filtering to collect the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a light yellow crystal, namely a formula I-2, wherein the yield is 81.4%, and the m.p.238-240 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.57(3H，t，CH₃)，2.24(3H，s，COCH₃) 2.83-3.56 (8H, m, piperazine-H), 3.87(3H, s, OCH)₃),4.36(2H，q，N-CH₂) 6.74-7.86 (6H, m, 8-H, Ph-H and 2 '-H), 8.26(1H, d, 5-H), 8.55(1H, d, 3' -H),8.87(1H, s, 2-H); MS (m/z): 478[ M + H]⁺Calculating (C)₂₇H₂₈FN₃O₄)：477.54。

Example 3

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- (3, 4-dioxocinnamoyl) -quinolin-4 (1H) -one (I-3) having the chemical formula:

namely, Ar in the formula I is 3,4- (dioxymethylene) phenyl.

The preparation method of the compound comprises the following steps: get 1-Ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-ethanone V1.1 g (3.0mmol) was dissolved in 20mL of anhydrous ethanol, and 3, 4-dioxytolualdehyde 0.53g (3.5mmol) and piperidine, a base catalyst (0.1mL) were added. And (3) carrying out reflux reaction on the mixed reactants for 20h, standing at room temperature, filtering to collect the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a light yellow crystal, namely a formula I-3, wherein the yield is 87.6%, and the m.p.246-248 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.62(3H，t，CH₃)，2.25(3H，s，COCH₃) 2.75 to 3.53(8H, m, piperazine-H), 4.42(2H, q, N-CH)₂)，6.17(2H，s，OCH₂O), 6.78-7.93 (5H, m, 8-H, Ph-H and 2 '-H), 8.31(1H, d, 5-H), 8.54(1H, d, 3' -H),8.87(1H, s, 2-H); MS (m/z): 492[ M + H]⁺Calculating (C)₂₇H₂₆FN₃O₅)：491.52。

Example 4

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- (3,4, 5-trimethoxycinnamoyl) -quinolin-4 (1H) -one (I-4) having the chemical formula:

namely, Ar in the formula I is 3,4, 5-trimethoxyphenyl.

The preparation method of the compound comprises the following steps: 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.63g (3.2mmol) of 3,4, 5-trioxybenzaldehyde and piperidine, a base catalyst (0.1mL) were added. And (3) carrying out reflux reaction on the mixed reactants for 20h, standing at room temperature, filtering to collect the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a light yellow crystal, namely a formula I-4, wherein the yield is 70.2%, and m.p.230-232 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.57(3H，t，CH₃)，2.25(3H，s，COCH₃) 2.78 to 3.56(8H, m, piperazine-H), 3.88, 3.95(9H, 2s, 3 XOCH)₃)，4.46(2H，q，N-CH₂) 6.68-7.87 (4H, m, 8-H, Ph-H and 2 '-H), 8.30(1H, d, 5-H), 8.53(1H, d, 3' -H),8.87(1H, s, 2-H); MS (m/z): 538[ M + H]⁺Calculating (C)₂₉H₃₂FN₃O₆)：537.59。

Example 5

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- (4-methylcinnamoyl) -quinolin-4 (1H) -one (I-5) having the chemical formula:

namely, Ar in the formula I is p-methyl-phenyl.

The preparation method of the compound comprises the following steps: 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.58g (4.8mmol) of 4-methylbenzaldehyde and piperidine, a basic catalyst (0.1mL) were added. And (3) carrying out reflux reaction on the mixed reactants for 15h, standing at room temperature, filtering to collect the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a light yellow crystal, namely a formula I-5, wherein the yield is 65.4%, and m.p.217-219 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.57(3H，t，CH₃)，2.26,2.32(6H，2s，Ph-CH₃And COCH₃) 2.70 to 3.53(8H, m, piperazine-H), 4.42(2H, q, N-CH)₂) 6.66-7.85 (6H, m, 8-H, Ph-H and 2 '-H), 8.28(1H, d, 5-H), 8.52(1H, d, 3' -H),8.83(1H, s, 2-H); MS (m/z): 462[ M + H ]]⁺Calculating (C)₂₇H₂₈FN₃O₃)：461.54。

Example 6

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- (4-fluorocinnamoyl) -quinolin-4 (1H) -one (I-6) having the chemical formula:

namely, Ar in the formula I is p-fluoro-phenyl.

The preparation method of the compound comprises the following steps: dissolving 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-ethanone in 20mL of anhydrous ethanol, and adding 4-fluorobenzaldehyde0.48g (3.8mmol) and piperidine (0.1mL) as the base catalyst. And (3) carrying out reflux reaction on the mixed reactants for 15h, standing at room temperature, filtering to collect the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a light yellow crystal, namely a formula I-6, wherein the yield is 74.8%, and the m.p. is 233-235 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.62(3H，t，CH₃)，2.30(3H，s，COCH₃) 2.85 to 3.63(8H, m, piperazine-H), 4.52(2H, q, N-CH)₂) 6.78-8.08 (6H, m, 8-H, Ph-H and 2 '-H), 8.36(1H, d, 5-H), 8.57(1H, d, 3' -H),8.90(1H, s, 2-H); MS (m/z): 466[ M + H]⁺Calculating (C)₂₆H₂₅F₂N₃O₃)：465.50。

Example 7

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- (4-chlorocinnamoyl) -quinolin-4 (1H) -one (I-7) having the chemical formula:

namely, Ar in the formula I is p-chlorophenyl.

The preparation method of the compound comprises the following steps: 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -1-yl-quinolin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.45g (3.2mmol) of 4-chlorobenzaldehyde and piperidine (0.1mL) as a base catalyst were added. And (3) refluxing and reacting the mixed reactants for 22h, standing at room temperature, filtering and collecting the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a light yellow crystal, namely a formula I-7, wherein the yield is 61.5%, and the m.p.221-223 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.60(3H，t，CH₃)，2.32(3H，s，COCH₃) 2.76 to 3.62(8H, m, piperazine-H), 4.48(2H, q, N-CH)₂) 6.76 to 8.05(6H, m, 8-H, Ph-H and 2 '-H), 8.33(1H, d, 5-H), 8.56(1H, d, 3' -H),8.92(1H, s, 2-H); MS (m/z): 482[ M + H ]]⁺(³⁵Cl), calculating (C)₂₆H₂₅FClN₃O₃)：481.96。

Example 8

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- (4-bromocinnamoyl) -quinolin-4 (1H) -one (I-8) having the chemical formula:

namely, Ar in the formula I is p-bromophenyl.

The preparation method of the compound comprises the following steps: 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.67g (3.6mmol) of 4-bromobenzaldehyde and piperidine (0.1mL) as a base catalyst were added. And (3) carrying out reflux reaction on the mixed reactants for 24 hours, standing at room temperature, filtering and collecting the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a light yellow crystal, namely a formula I-8, wherein the yield is 70.1%, and the m.p.238-240 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.62(3H，t，CH₃)，2.28(3H，s，COCH₃) 2.76 to 3.57(8H, m, piperazine-H), 4.47(2H, q, N-CH)₂) 6.82-8.04 (6H, m, 8-H, Ph-H and 2 '-H), 8.36(1H, d, 5-H), 8.55(1H, d, 3' -H),8.89(1H, s, 2-H); MS (m/z): 526 and 528[ M + H ]]⁺(⁷⁹Br and⁸¹br), calculating (C)₂₆H₂₅FBrN₃O₃)：526.41。

Example 9

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- (4-nitrocinnamoyl) -quinolin-4 (1H) -one (I-9) having the chemical formula:

namely, Ar in the formula I is p-nitrophenyl.

The preparation method of the compound comprises the following steps: 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.54g (3.6mmol) of 4-nitrobenzaldehyde and piperidine, a base catalyst (0.1mL) were added. And (3) carrying out reflux reaction on the mixed reactants for 24 hours, standing at room temperature, filtering and collecting the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a yellow crystal, namely a formula I-9, wherein the yield is 73.5%, and the m.p.243-246 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.66(3H，t，CH₃)，2.32(3H，s，COCH₃) 2.86 to 3.67(8H, m, piperazine-H), 4.54(2H, q, N-CH)₂) 6.96-7.93 (2H, m, 8-H and 2 '-H), 8.44-8.68 (6H, 5-H, 3' -H and Ph-H),9.07(1H, s, 2-H); MS (m/z): 493 calculating (C)₂₆H₂₅FN₄O₅)：492.51。

Example 10

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- (4-hydroxy-cinnamoyl) -quinolin-4 (1H) -one (I-10) having the chemical formula:

namely, Ar in the formula I is 4-hydroxy-phenyl.

The preparation method of the compound comprises the following steps: 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.49g (4.0mmol) of 4-hydroxy-benzaldehyde and piperidine, a base catalyst (0.1mL) were added. And (3) carrying out reflux reaction on the mixed reactants for 20h, standing at room temperature, filtering to collect the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a yellow crystal, namely a formula I-10, wherein the yield is 58.6%, and the m.p.230-232 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.57(3H，t，CH₃)，2.26(3H，s，COCH₃) 2.68 to 3.56(8H, m, piperazine-H), 4.45(2H, q, N-CH)₂) 6.86-7.85 (6H, m, 8-H, Ph-H and 2 '-H), 8.32(1H, d, 5-H), 8.48(1H, d, 3' -H),8.87(1H, s,2-H), 10.55(1H, s, OH); MS (m/z): 464, calculating (C)₂₅H₂₄FN₃O₄)：463.51。

Example 11

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- [3- (pyridin-3-yl) acryloyl ] -quinolin-4 (1H) -one (I-11) having the chemical formula:

namely, Ar in the formula I is 3-pyridyl.

The preparation method of the compound comprises the following steps: 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.37g (3.6mmol) of 3-pyridylaldehyde and piperidine, a basic catalyst (0.1mL) were added. And (3) carrying out reflux reaction on the mixed reactants for 16h, standing at room temperature, filtering to collect the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a yellow crystal, namely a formula I-11, wherein the yield is 80.4%, and the m.p.241-243 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.67(3H，t，CH₃)，2.33(3H，s，COCH₃) 3.07 to 3.68(8H, m, piperazine-H), 4.64(2H, q, N-CH)₂) 7.18-7.96 (2H, m, 8-H and 2 '-H), 8.34-8.91 (6H, 5-H, 3' -H and pyridine-H), 9.15(1H, s, 2-H); MS (m/z): 449, calculating (C)₂₅H₂₅FN₄O₃)：448.50。

Example 12

1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -3- [3- (furan-2-yl) acryloyl ] quinolin-4 (1H) -one (I-12) having the chemical formula:

namely, Ar in the formula I is 2-furyl.

The preparation method of the compound comprises the following steps: the preparation method of the compound comprises the following steps: 1.1g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-acetylpiperazin-1-yl) -quinolin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.38g (4.0mmol) of 2-furfural and piperidine (0.1mL) as a base catalyst were added. And (3) refluxing and reacting the mixed reactants for 18h, standing at room temperature, filtering and collecting generated solid, and recrystallizing with absolute ethyl alcohol to obtain a yellow crystal, namely a formula I-12, wherein the yield is 63.7%, and the m.p.232-234 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.58(3H，t，CH₃)，2.30(3H，s，COCH₃) 2.76 to 3.63(8H, m, (piperazine-H), 4.58(2H, q, N-CH)₂) 6.87-7.88 (5H, m, 8-H, 2 '-H and furan-H), 8.45(1H, d, 5-H), 8.57(1H, d, 3' -H),8.88(1H, s, 2-H); MS (m/z):438[M+H]⁺calculating (C)₂₄H₂₄FN₃O₄)：437.47。

Test examples

One, embodiment 1-12 provides a N-acetyl norfloxacin propenone derivatives in vitro antitumor activity determination

1. Test sample

15 of the propenone derivatives of N-acetyl norfloxacin provided in examples 1-12, as well as the classical antitumor TOPO inhibitor 10-Hydroxycamptothecin (HC), the chalcone tyrosinase inhibitor Sunitinib (SN), the broad spectrum anticancer drug Doxorubicin (DOX), and the parent compound N-Acetyl Norfloxacin (ANF) were used as test samples, wherein HC, SN, and ANF were used as control groups, and the samples of examples 1-12 were used as experimental groups;

thiazole blue (MTT), HC, SN and ANF are all products of Sigma company; the RPMI-1640 culture solution is a product of GIBCO company; other used reagents are all domestic analytical pure reagents.

The experimental cancer cell strains are respectively a human non-small cell lung cancer cell strain A549, a human kidney cancer cell strain 769-P, a human hepatoma cell strain Hep-3B, a human gastric cancer cell strain HGC27, a human pancreatic cancer cell strain Panc-1 and a human leukemia cell strain HL60, which are purchased from Shanghai cell banks of Chinese academy of sciences. The human renal clear cell carcinoma cell sunitinib-resistant strain 7SuR was purchased from shanghai zel biotechnology limited, and the normal cell was obtained from african green monkey kidney cell line VERO and purchased from shanghai tong biology limited.

2. Measurement method

The determination method comprises the following specific steps:

1) firstly, the 15 samples were dissolved in dimethyl sulfoxide (DMSO) to prepare 1.0X 10^{- 4}The stock solution with mol.L-1 concentration is diluted into a stock solution with 5 concentration gradients (0.1, 1.0, 5.0, 10.0, 50.0 mu mol.L) by using a calf serum RPMI-1640 culture solution with a mass percent concentration of 10 percent^-1) The working fluid of (1);

2) collecting non-small cell lung cancer cell strain A549, human renal cancer cell strain 769-P, human liver cancer cell strain Hep-3B, and human gastric cancer in logarithmic growth phaseCell line HGC27, human pancreatic cancer cell line Panc-1 and human leukemia cell line HL60, human renal clear cell carcinoma cell sunitinib drug-resistant line 7SuR and African green monkey kidney cell line VERO, 6000 cells per well were inoculated into 96-well plate, then working solution with 5 concentration gradients was added to each of the 15 samples, and after 48 hours, 5 g.L/well was added^–1mu.L of MTT (thiazole blue) solution was added, and after further culturing for 4 hours, 100. mu.L of a 10% by mass Sodium Dodecyl Sulfate (SDS) solution was added. Culturing for 24 hours, and then measuring an absorbance (OD) value at a wavelength of 570nm by using a microplate reader;

3) the inhibition rate of the test samples with different concentrations on the cancer cells is calculated according to the following formula:

cancer cell inhibition rate ═ [ (1-experimental OD value)/control OD value ] × 100%;

then, linear regression is carried out on the cancer cell inhibition rate corresponding to each concentration by the pair value of each concentration of the test sample to obtain a dose-effect equation, and the half Inhibition Concentration (IC) of the test sample to the experimental cancer cell is calculated from the obtained dose-effect equation₅₀) (ii) a Each data was measured in triplicate and averaged, the results are shown in Table 1.

TABLE 1 antitumor Activity (IC) of the test samples₅₀)

As can be seen from Table 1, the inhibitory activity of the compounds provided in examples 1-12 on 7 cancer cells of experiment is significantly stronger than that of the parent compound N-acetyl norfloxacin, especially the growth inhibitory activity of some compounds on human non-small cell lung cancer cell line A549 is stronger than that of the control Hydroxycamptothecin (HC), tyrosine kinase inhibitors Sunitinib (SN) and adriamycin (DOX), and the IC is that₅₀The value is close to nanomolar concentration, and the method has potential new drug research value. More significantly, the compounds provided in examples 1 to 12 also show extremely strong sensitivity to sunitinib-resistant strain 7SuR, and show strong resistance to drugs; simultaneously has low toxicity to normal cell VERO and has patent medicineThe nature of sex. Therefore, according to the general approach of drug development, the conventional antitumor in vitro screening is carried out, and then the targeted research is carried out, so that the compound has strong antitumor activity, drug resistance activity and lower cytotoxicity, and can be used for preparing antitumor drugs by salifying with acid acceptable for human bodies or mixing with medicinal carriers.