阅读说明：本技术 一种n-甲基依诺沙星的丙烯酮衍生物及其制备方法和应用 (N-methylenoxacin acrylketone derivative and preparation method and application thereof ) 是由 马云峰 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-甲基依诺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-甲基依诺沙星的丙烯酮衍生物,实现了氟萘啶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-methylenoxacin, a preparation method and application thereof, wherein the acrylic ketone derivative adopts a chemical structure general formula as shown in the following formula I:)

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

2. the preparation method of the propenone derivative of N-methylenoxacin according to claim 1, characterized by comprising the following steps:

1) taking N-methylenoxacin shown in a formula II as a raw material, reacting with Carbonyldiimidazole (CDI) to prepare an N-methylenoxacin 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-methylenoxacin shown in a formula IV; finally, the N-methyl enoxacin C-3 ethanone shown in the formula V is prepared by the hydrolysis decarboxylation reaction of the formula IV:

2) the propenone derivative of N-methylenoxacin as shown in claim 1 can be prepared by carrying out Claisen-Schmidt condensation reaction on N-methylenoxacin C-3 ethanone shown in 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 propenone derivative of N-methylenoxacin according to claim 2, characterized in that the molar ratio of N-methylenoxacin represented by formula II to CDI is 1: 1.0-2.0, the molar ratio of N-methylenoxacin imidazole amide represented by formula III to monoethyl malonate potassium salt is 1: 1.0-1.5, and the molar ratio of N-methylenoxacin-3 ethanone represented by formula V to aromatic aldehyde is 1: 1.0-2.0.

4. The use of the propenone derivative of N-methylenoxacin according to claim 1 for the preparation of an anti-tumor medicament.

5. The application of the propenone derivative of N-methylenoxacin in the preparation of the antitumor drug according to claim 4, is characterized in that 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 medicine synthesis, and particularly relates to an acrylketone derivative of N-methylenoxacin, a preparation method of the acrylketone derivative of the N-methylenoxacin, and application of the acrylketone derivative in an antitumor medicine.

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 C-3 carboxyl of fluoronaphthyridone by 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 bioactivity of the chalcone, the invention uses the skeleton of the dominant pharmacophore of the fluoroquinolone medicament N-methylenoxacin, namely '1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -naphthyridin-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 an acrylketone derivative of N-methylenoxacin, which has the function and the efficacy of resisting tumors, and also provides a preparation method of the acrylketone derivative of the N-methylenoxacin.

In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: the N-methylenoxacin acrylketone derivative 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-methylenoxacin is characterized in that N-methylenoxacin shown in a formula II is commercially obtained and is prepared by taking N-methylenoxacin as a raw material;

the preparation method comprises the following specific steps:

1) the preparation method of the enoxacin imidazole amide compound shown in the formula III is that N-methyl enoxacin shown in the formula II is used as a raw material and reacts with Carbonyldiimidazole (CDI), and the preparation method comprises the following steps:

taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]20g (60.0mmol) of naphthyridine-4 (1H) -ketone-3-carboxylic acid II is dissolved in 500mL of anhydrous acetonitrile, 15.2g (94.0mmol) of carbonyl diimidazole is added, and the mixed reactants are stirred in a water bath and refluxed until the raw 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 with the yield of 81.4 percent and the m.p.234-236 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.57(3H，t，CH₃)，2.36(3H，s，CH₃) 3.15 to 3.57(8H, m, piperazine-H), 4.68(2H, q, N-CH)₂) 7.06-7.36 (2H, m, imidazole-H), 8.25(1H, d, 5-H), 8.28(1H, s, imidazole-H), 8.96(1H, s, 2-H); MS (m/z): 385[ M + H ]]⁺Calculating (C)₁₉H₂₁FN₆O₂)：384.42。

As a further improvement, the molar ratio of the N-methylenoxacin 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-methylenoxacin 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 a C-3 formyl ethyl acetate compound of N-methylenoxacin shown in a formula IV, and the specific preparation method is as follows:

taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- (1H-imidazole-1-formyl) - [1, 8%]Naphthyridine-4 (1H) -ketone formula III 15g (39.0mmol), magnesium chloride 6.6g (69.1mmol) and malonic acid monoethyl ester potassium salt 8.3g (49.0mmol) are sequentially added into 600mL of anhydrous acetonitrile, triethylamine 12.2g (12.0mmol) is added dropwise under stirring in an ice bath, and the mixed reactant is stirred in a water bath and refluxed until the raw material III disappears. The solvent was evaporated under reduced pressure, 500mL of water was added, the mixture was extracted with methylene chloride (3X 150mL), the organic phases were combined, washed with water (3X 200mL), washed with saturated brine (2X 150mL), and dried over anhydrous sodium sulfate. And recovering dichloromethane at normal pressure, and recrystallizing the residue with absolute ethyl alcohol to obtain an off-white crystal shown as a formula IV, wherein the yield is 75.3%, and m.p.220-222 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.32,1.63(6H，2t，2×CH₃)，2.38(3H，s，N-CH₃) 3.17 to 3.60(8H, m, piperazine-H), 4.18(2H, s, COCH)₂CO)，4.28(2H，q，CO₂CH₂)，4.70(2H，q，N-CH₂)，8.26(1H，d，5-H)，8.97(1H，s,2-H)；MS(m/z)：405[M+H]⁺Calculating (C)₂₀H₂₅FN₄O₄)：404.45。

3) The C-3 ethyl formylacetate compound of the N-methylenoxacin shown in the formula IV is subjected to hydrolysis decarboxylation reaction by using a sodium hydroxide aqueous solution with the mass fraction of 6 percent, so that the C-3 ethanone compound of the N-methylenoxacin shown in the formula V can be conveniently prepared, and the specific preparation method is as follows:

taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]Naphthyridin-4 (1H) -one-3-formylacetic acid ethyl ester formula IV 10g (25).0mmol) of the crude product is suspended in 200mL of 6 percent sodium hydroxide aqueous solution, and the mixture is stirred and refluxed in an oil bath 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 75.6%, and m.p.234-236 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.63(3H，t，CH₃)，2.38(3H，s，N-CH₃)，2.44(3H，s，COCH₃) 3.07 to 3.63(8H, m, piperazine-H), 4.67(2H, q, N-CH)₂)，8.24(1H，d，5-H)，8.96(1H，s,2-H)；MS(m/z)：333[M+H]⁺Calculating (C)₁₇H₂₁FN₄O₂)：332.38。

4) C-3 ethanone of N-methylenoxacin shown in the 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 through treatment, wherein the target compound is shown in the 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: 1.0g (3.0mmol) of 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1,8] naphthyridin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and an aromatic aldehyde (3.0mmol) and a 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-methylenoxacin 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-methylenoxacin in preparing the 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-methylenoxacin is designed and synthesized by effectively combining a fluoroquinolone framework and an aryl acrylketone pharmacophore based on the splicing principle of the pharmacophores, realizes the complementation and the activity superposition of the pharmacophores with different structures, thereby achieving the effects of synergism, toxicity reduction and drug resistance, and can be developed as an anti-tumor drug with a brand new structure.

Detailed Description

Example 1

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3-cinnamoyl- [1,8] naphthyridin-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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]1.0g (3.0mmol) of naphthyridin-4 (1H) -one-3-ethanone V is dissolved in 20mL of absolute ethanol, and 0.40g (3.8mmol) of benzaldehyde and a base catalyst piperidine (0.1mL) are 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 68.2%, and the m.p.225-227 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.64(3H，t，CH₃)，2.38(3H，s，N-CH₃) 3.15 to 3.67(8H, m, piperazine-H), 4.67(2H, q, N-CH)₂) 7.36-7.87 (6H, m, Ph-H and 2 '-H), 8.28(1H, d, 5-H), 8.57(1H, d, 3' -H),9.06(1H, s, 2-H); MS (m/z): 421[ M + H]⁺Calculating (C)₂₄H₂₅FN₄O₂)：420.49。

Example 2

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- (4-methoxycinnamoyl) - [1,8] naphthyridin-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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]1.0g (3.0mmol) of naphthyridin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.57g (4.2mmol) of 4-methoxybenzaldehyde and piperidine (0.1mL) as a base catalyst were added. And (3) refluxing and reacting the mixed reactants for 20 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-2, wherein the yield is 73.6%, and the m.p.234-236 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.66(3H，t，CH₃)，2.42(3H，s，N-CH₃) 3.14 to 3.65(8H, m, piperazine-H), 3.90(3H, s, OCH)₃),4.68(2H，q，N-CH₂) 7.38-7.84 (5H, m, Ph-H and 2 '-H), 8.33(1H, d, 5-H), 8.60(1H, d, 3' -H),9.10(1H, s, 2-H); MS (m/z): 451[ M + H ]]⁺Calculating (C)₂₅H₂₇FN₄O₃)：450.52。

Example 3

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- (3, 4-dioxocinnamoyl) - [1,8] naphthyridin-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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]1.0g (3.0mmol) of naphthyridin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.53g (3.5mmol) of 3, 4-dioxytolualdehyde and piperidine (0.1mL) as a base catalyst were added. Refluxing the mixed reactants for 20h, standing at room temperature, filtering to collect the generated solid, and recrystallizing with anhydrous ethanol to obtain light yellow crystal of formula I-3 with yield of 80.3%, m.p.238～240℃。¹H NMR(400MHz,CD₃Cl)δ：1.66(3H，t，CH₃)，2.42(3H，s，N-CH₃) 3.15 to 3.67(8H, m, piperazine-H), 4.68(2H, q, N-CH)₂)，6.23(2H，s，OCH₂O), 7.40-7.87 (4H, m, Ph-H and 2 '-H), 8.35(1H, d, 5-H), 8.58(1H, d, 3' -H),9.16(1H, s, 2-H); MS (m/z): 465[ M + H]⁺Calculating (C)₂₅H₂₅FN₄O₄)：464.50。

Example 4

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- (3,4, 5-trimethoxycinnamoyl) - [1,8] naphthyridin-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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]1.0g (3.0mmol) of naphthyridin-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 (0.1mL) as a basic catalyst 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 73.5%, and the m.p.231-233 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.65(3H，t，CH₃)，2.38(3H，s，N-CH₃) 3.13 to 3.66(8H, m, piperazine-H), 3.87, 3.95(9H, 2s, 3 XOCH)₃)，4.68(2H，q，N-CH₂) 7.42-7.86 (3H, m, Ph-H and 2 '-H), 8.36(1H, d, 5-H), 8.57(1H, d, 3' -H),9.15(1H, s, 2-H); MS (m/z): 511[ M + H]⁺Calculating (C)₂₇H₃₁FN₄O₅)：510.57。

Example 5

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- (4-methylcinnamoyl) - [1,8] naphthyridin-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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]1.0g (3.0mmol) of naphthyridin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.58g (4.8mmol) of 4-methylbenzaldehyde and piperidine (0.1mL) as a base catalyst 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 62.8%, and the m.p.216-218 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.62(3H，t，CH₃)，2.30(3H，s，Ph-CH₃)，2.38(3H，s，N-CH₃) 3.07 to 3.63(8H, m, piperazine-H), 4.66(2H, q, N-CH)₂) 7.38-7.86 (5H, m, Ph-H and 2 '-H), 8.32(1H, d, 5-H), 8.54(1H, d, 3' -H),9.12(1H, s, 2-H); MS (m/z): 435[ M + H ]]⁺Calculating (C)₂₅H₂₇FN₄O₂)：434.52。

Example 6

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- (4-fluorocinnamoyl) - [1,8] naphthyridin-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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]1.0g (3.0mmol) of naphthyridin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.48g (3.8mmol) of 4-fluorobenzaldehyde and piperidine (0.1mL) as a base catalyst 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-6, wherein the yield is 77.3%, and m.p.231-233 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.68(3H，t，CH₃)，2.42(3H，s，N-CH₃)，3.17～3.72(8H，m，piperazine-H), 4.73(2H, q, N-CH)₂) 7.44-8.08 (5H, m, Ph-H and 2 '-H), 8.36(1H, d, 5-H), 8.62(1H, d, 3' -H),9.18(1H, s, 2-H); MS (m/z): 439[ M + H]⁺Calculating (C)₂₄H₂₄F₂N₄O₂)：438.48。

Example 7

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- (4-chlorocinnamyl) - [1,8] naphthyridin-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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -1-yl- [1,8]1.0g (3.0mmol) of naphthyridin-4 (1H) -one-3-ethanone V is dissolved in 20mL of absolute ethanol, and 0.45g (3.2mmol) of 4-chlorobenzaldehyde and a base catalyst piperidine (0.1mL) are 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-7, wherein the yield is 58.4%, and the m.p.221-223 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.66(3H，t，CH₃)，2.41(3H，s，N-CH₃) 3.15 to 3.68(8H, m, (piperazine-H), 4.67(2H, q, N-CH)₂) 7.43-8.06 (5H, m, Ph-H and 2 '-H), 8.36(1H, d, 5-H), 8.60(1H, d, 3' -H),9.16(1H, s, 2-H); MS (m/z): 455[ M + H]⁺(³⁵Cl), calculating (C)₂₄H₂₄FClN₄O₂)：454.94。

Example 8

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- (4-bromocinnamoyl) - [1,8] naphthyridin-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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]1.0g (3.0mmol) of naphthyridin-4 (1H) -one-3-ethanone V is dissolved in 20mL of anhydrous ethanol, and 0.67g (3.6mmol) of 4-bromobenzaldehyde and piperidine (0.1mL) as a base catalyst are added. And (3) refluxing and reacting 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 73.8%, and the m.p.234-236 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.66(3H，t，CH₃)，2.43(3H，s，N-CH₃) 3.15 to 3.68(8H, m, piperazine-H), 4.67(2H, q, N-CH)₂) 7.40-8.06 (5H, m, Ph-H and 2 '-H), 8.36(1H, d, 5-H), 8.62(1H, d, 3' -H),9.17(1H, s, 2-H); MS (m/z): 499 and 501[ M + H [ ]]⁺(⁷⁹Br and⁸¹br), calculating (C)₂₄H₂₄FBrN₄O₂)：498.39。

Example 9

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- (4-nitrocinnamoyl) - [1,8] naphthyridin-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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]1.0g (3.0mmol) of naphthyridin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.54g (3.6mmol) of 4-nitrobenzaldehyde 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 to collect the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a yellow crystal, namely the formula I-9, wherein the yield is 57.4%, and the m.p.243-245 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.74(3H，t，CH₃)，2.46(3H，s，N-CH₃) 3.30 to 3.75(8H, m, piperazine-H), 4.74(2H, q, N-CH)₂) 7.46(1H, d, 2 '-H), 8.46-8.68 (5H, 3' -H and Ph-H),9.22(1H, s, 2-H); MS (m/z): 466, calculating (C)₂₄H₂₄FN₅O₄)：465.49。

Example 10

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- (4-hydroxy-cinnamoyl) - [1,8] naphthyridin-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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]1.0g (3.0mmol) of naphthyridin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.49g (4.0mmol) of 4-hydroxy-benzaldehyde and piperidine (0.1mL) as a base catalyst 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 61.8.0%, and m.p.232-235 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.64(3H，t，CH₃)，2.37(3H，s，N-CH₃) 3.13 to 3.64(8H, m, piperazine-H), 4.65(2H, q, N-CH)₂) 7.38-7.86 (5H, m, Ph-H and 2 '-H), 8.36(1H, d, 5-H), 8.57(1H, d, 3' -H),8.96(1H, s,2-H), 10.67(1H, s, OH); MS (m/z): 437, calculating (C)₂₄H₂₅FN₄O₃)：436.49。

Example 11

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- [3- (pyridin-3-yl) acryloyl ] - [1,8] naphthyridin-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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]1.0g (3.0mmol) of naphthyridin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.37g (3.6mmol) of 3-pyridylaldehyde and piperidine (0.1mL) as a base catalyst were added. The mixed reactants are refluxed and reacted for 15 hours, placed at room temperature and filteredAnd recrystallizing the generated solid with absolute ethyl alcohol to obtain a yellow crystal substance shown as the formula I-11, wherein the yield is 82.0%, and the m.p.237-239 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.72(3H，t，CH₃)，2.44(3H，s，N-CH₃) 3.23 to 3.76(8H, m, piperazine-H), 4.72(2H, q, N-CH)₂) 7.47(1H, d, 2 '-H), 8.42-8.96 (6H, 5-H, 3' -H and pyridine-H), 9.18(1H, s, 2-H); MS (m/z): 422, calculating (C)₂₃H₂₄FN₅O₂)：421.48。

Example 12

1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) -3- [3- (furan-2-yl) acryloyl ] [1,8] naphthyridin-4 (1H) -one (I-12) having the chemical structure:

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: taking 1-ethyl-6-fluoro-7- (4-methylpiperazin-1-yl) - [1, 8%]1.0g (3.0mmol) of naphthyridin-4 (1H) -one-3-ethanone V was dissolved in 20mL of anhydrous ethanol, and 0.38g (4.0mmol) of 2-furan aldehyde 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 the generated solid, and recrystallizing with absolute ethyl alcohol to obtain a yellow crystal, namely a formula I-12, wherein the yield is 62.4%, and the m.p. is 233-235 ℃.¹H NMR(400MHz,CD₃Cl)δ：1.65(3H，t，CH₃)，2.42(3H，s，N-CH₃) 3.15 to 3.68(8H, m, piperazine-H), 4.67(2H, q, N-CH)₂) 7.23-7.84 (4H, m, 2 '-H and furan-H), 8.45(1H, d, 5-H), 8.63(1H, d, 3' -H),9.16(1H, s, 2-H); MS (m/z): 411[ M + H]⁺Calculating (C)₂₂H₂₃FN₄O₃)：410.45。

Test examples

One, embodiment 1-12 provides an in vitro antitumor activity assay of acrylketone derivatives of N-methylenoxacin

1. Test sample

15 samples of the propenone derivative of N-methylenoxacin provided in examples 1 to 12, a classical antitumor TOPO inhibitor 10-Hydroxycamptothecin (HC), a chalcone tyrosinase inhibitor Sunitinib (SN), a broad-spectrum anticancer drug Doxorubicin (DOX) and a parent compound N-Methylenoxacin (MEN) are taken as test samples, wherein HC, SN and MEN are taken as control experimental groups, and examples 1 to 12 are taken as test experimental groups;

thiazole blue (MTT), HC, SN and MEN 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}mol·L^-1Stock solution of concentration, then diluting the stock solution with 10% calf serum RPMI-1640 culture solution to have 5 concentration gradients (0.1, 1.0, 5.0, 10.0, 50.0 μmol. L)^-1) The working fluid of (1);

2) taking non-small cell lung cancer cell strain A549, human kidney cancer cell strain 769-P, human liver cancer cell strain Hep-3B, human gastric cancer cell strain HGC27, human pancreatic cancer cell strain Panc-1, human leukemia cell strain HL60, human renal clear cell cancer cell sunitinib drug-resistant strain 7SuR and African green monkey kidney cell strain VERO in logarithmic growth phase, inoculating 6000 cells in each hole to a 96-hole plate, then respectively adding working solution with 5 concentration gradients of the 15 samples, and adding 5 g.L.in each hole after 48 hours^–1MTT (thiazole blue) solution 10. mu.L, continue to get againAfter 4 hours of incubation, 100. mu.L of a 10% strength 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 the experiment is significantly stronger than that of the parent compound N-methylenoxacin, 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), the tyrosine kinase inhibitors Sunitinib (SN) and adriamycin (DOX), and the IC of the compounds is₅₀The value reaches nanomolar concentration, and the method has the value of new drug development. More significantly, the compounds provided in examples 1 to 12 also show extremely strong sensitivity to sunitinib-resistant strain 7SuR, show strong drug-resistant activity, and simultaneously show low toxicity to normal cells VERO, and have the property of becoming drug-resistant. 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.