A benzonaphthyridin-5-one derivative, its preparation method and application

Abstract

The present invention provides a benzonaphthyridin-5-one derivative, a preparation method thereof and an application, relating to biomedicine. The benzonaphthyridin-5-one derivative provided by the present invention is a class of compounds with unique structures, having ROCK kinase inhibitory activity, and exerting a significant anti-tumor effect by inhibiting the proliferation, migration and invasion of tumor cells and improving the tumor microenvironment. The benzonaphthyridin-5-one derivative provided by the present invention shows important application potential in the fields of drug development, new material design and organic electronic devices. Especially in medicinal chemistry, it is expected to become the core skeleton of novel anti-cancer, antibacterial, antiviral and anti-inflammatory drugs. Due to the complex structure of the benzonaphthyridin-5-one derivative, its efficient synthesis has always faced challenges. The preparation method of the benzonaphthyridin-5-one derivative provided by the present invention has simple process, high synthesis efficiency and low production cost, and is suitable for industrial production.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly relates to a benzonaphthyridin-5-one derivative, a preparation method thereof, and an application thereof. Background Art

[0002] Benzonaphthyridin-5-one derivatives are a class of compounds with a polycyclic aromatic structure. The core structure includes a benzene ring and a naphthyridinone structure containing nitrogen. They have unique physical and chemical properties and a wide range of biological activities, showing significant potential application values in anti-cancer, anti-tumor, anti-inflammatory, antibacterial, antiviral, and neuroprotection aspects. For example, the compounds shown in Formulas 1 to 2 both have ROCK kinase inhibitory activity. Among them, the compound shown in Formula 2 is a small molecule amyloid protein regulator, which can increase the levels of synaptophysin and synaptopodin, and play a significant anti-tumor effect by inhibiting the proliferation, migration, and invasion of tumor cells and improving the tumor microenvironment. However, the existing types of compounds with ROCK kinase inhibitor activity are relatively few, which limits their applications.

[0003] Summary of the Invention

[0004] In view of this, the purpose of the present invention is to provide a benzonaphthyridin-5-one derivative, a preparation method thereof, and an application thereof. The benzonaphthyridin-5-one derivative provided by the present invention has ROCK kinase inhibitory activity.

[0005] To achieve the above-mentioned invention purpose, the present invention provides the following technical solutions:

[0006] The present invention provides a benzonaphthyridin-5-one derivative, which is characterized by having the structure shown in Formula I:

[0007]

[0008] Wherein, R 1 includes hydrogen, methyl, methoxy, or chlorine; R 2 includes benzyl, methyl, or ethyl; R 3 includes hydrogen, methyl, methoxy, or chlorine, and the number of R 3 on a single benzene ring is 1 to 2.

[0009] Preferably, the benzonaphthyridin-5-one derivative has the structure shown in Formula 3aa, Formula 3ba, Formula 3ca, Formula 3da, Formula 3ea, Formula 3ab, Formula 3ac, Formula 3ad, or Formula 3ae:

[0010]

[0011] The present invention provides a preparation method of the benzonaphthyridin-5-one derivative according to the above technical solution, including the following steps:

[0012] The ring-expansion reaction of Compound 1 and Compound 2 is carried out in the presence of an organometallic catalyst to obtain the benzonaphthyridin-5-one derivative;

[0013]

[0014] Preferably, the organometallic catalyst includes one or more of dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer, dichloro(pentamethylcyclopentadienyl)ruthenium(III) polymer, and bis[2-(2,4-difluorophenyl)-5-trifluoromethylpyridine][2-2'-bi(4-tert-butylpyridine)]iridium bis(hexafluorophosphate).

[0015] Preferably, the molar ratio of Compound 1 to the organometallic catalyst is 1:0.01 - 0.05.

[0016] Preferably, the ring-expansion reaction is carried out in the presence of an additive, and the additive includes cuprous salt and / or cuprous oxide.

[0017] Preferably, the molar ratio of Compound 1 to the additive is 1:0.5 - 2.5.

[0018] Preferably, the molar ratio of Compound 1 to Compound 2 is 1:1.25 - 5.

[0019] Preferably, the temperature of the ring-expansion reaction is 80 - 150 °C, and the time is 10 - 20 h;

[0020] The organic solvents used in the ring-expansion reaction include chloroalkanes and / or ester solvents;

[0021] The ring-expansion reaction is carried out under a protective atmosphere.

[0022] The present invention also provides the application of the benzonaphthyridin-5-one derivative described in the above technical solution in the preparation of anti-tumor drugs, anti-inflammatory drugs, antibacterial drugs, antiviral drugs, neuroprotective drugs, drugs for reducing amyloid burden, drugs for treating cardiovascular diseases, or organic electronic devices.

[0023] The benzonaphthyridin-5-one derivative provided by the present invention is a class of compounds with unique structures, having ROCK kinase inhibitory activity. By inhibiting the proliferation, migration, and invasion of tumor cells and improving the tumor microenvironment, it exerts a significant anti-tumor effect. The benzonaphthyridin-5-one derivative provided by the present invention shows important application potential in the fields of drug development, new material design, and organic electronic devices. Especially in medicinal chemistry, it is expected to become the core skeleton of novel anti-cancer, antibacterial, antiviral, and anti-inflammatory drugs.

[0024] Due to the complex structure of benzonaphthyridin-5-one derivatives, their efficient synthesis has always been a challenge. The preparation method of benzonaphthyridin-5-one derivatives provided by the present invention has a simple process, is easy to operate, has high synthesis efficiency, the raw materials are cheap and easily available, the production cost is low, and it is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 1H NMR spectrum of compound 3aa;

[0026] Figure 2 13C NMR spectrum of compound 3aa;

[0027] Figure 3 1H NMR spectrum of compound 3ba;

[0028] Figure 4 13C NMR spectrum of compound 3ba;

[0029] Figure 5 1H NMR spectrum of compound 3ca;

[0030] Figure 6 13C NMR spectrum of compound 3ca;

[0031] Figure 7 1H NMR spectrum of compound 3da;

[0032] Figure 8 13C NMR spectrum of compound 3da;

[0033] Figure 9 1H NMR spectrum of compound 3ea;

[0034] Figure 10 13C NMR spectrum of compound 3ea;

[0035] Figure 11 1H NMR spectrum of compound 3ab;

[0036] Figure 12 13C NMR spectrum of compound 3ab;

[0037] Figure 13 1H NMR spectrum of compound 3ac;

[0038] Figure 14 13C NMR spectrum of compound 3ac;

[0039] Figure 15 1H NMR spectrum of compound 3ad;

[0040] Figure 16 13C NMR spectrum of compound 3ad;

[0041] Figure 17 1H NMR spectrum of compound 3ae;

[0042] Figure 18 This is the carbon spectrum of compound 3ae. Detailed implementation mode

[0043] The present invention provides a benzonaphthyridin-5-one derivative, which is characterized by having the structure shown in formula I:

[0044]

[0045] Wherein, R 1 includes hydrogen, methyl, methoxy or chlorine; R 2 includes benzyl, methyl or ethyl; R 3 includes hydrogen, methyl, methoxy or chlorine, and the number of R 3 on a single benzene ring is preferably 1 to 2.

[0046] In the present invention, the benzonaphthyridin-5-one derivative preferably has the structure shown in formula 3aa, formula 3ba, formula 3ca, formula 3da, formula 3ea, formula 3ab, formula 3ac, formula 3ad or formula 3ae:

[0047]

[0048] The present invention also provides a preparation method of the benzonaphthyridin-5-one derivative described in the above technical solution, including the following steps:

[0049] Perform a ring-expansion reaction on compound 1 and compound 2 in the presence of an organometallic catalyst to obtain the benzonaphthyridin-5-one derivative;

[0050]

[0051] Unless otherwise specified, the materials and equipment used in the present invention are all commercially available products in the art.

[0052] In the present invention, the molar ratio of compound 1 to compound 2 is preferably 1:1.25 to 5, and in specific examples, it can be 1:1.25, 1:1.5, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5 or 1:5.

[0053] In the present invention, the organometallic catalyst preferably comprises one or more of dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer, dichloro(pentamethylcyclopentadienyl)ruthenium(III) polymer, and bis[2-(2,4-difluorophenyl)-5-trifluoromethylpyridine][2-2'-bi(4-tert-butylpyridine)]iridium bis(hexafluorophosphate). When the organometallic catalyst is a mixture of two or more, there is no special limitation on the mass ratio of different organometallic catalysts in the present invention, and any ratio is acceptable. In the present invention, the molar ratio of Compound 1 to the organometallic catalyst is preferably 1:0.01 to 0.05, and in specific embodiments, it can be 1:0.01, 1:0.02, 1:0.03, 1:0.04, or 1:0.05.

[0054] In the present invention, the ring-expansion reaction is preferably carried out in the presence of an additive. The additive preferably comprises a cuprous salt and / or cuprous oxide; the cuprous salt preferably comprises cuprous halide, and more preferably comprises cuprous chloride and / or cuprous bromide. In the present invention, the molar ratio of Compound 1 to the additive is preferably 1:0.5 to 2.5, and in specific embodiments, it can be 1:0.5, 1:1, 1:1.5, 1:2, or 1:2.5. In the present invention, the use of the additive can improve the product yield.

[0055] In the present invention, the organic solvent used in the ring-expansion reaction preferably comprises a chloroalkane and / or an ester solvent, and more preferably a mixed solvent of a chloroalkane and an ester solvent; the chloroalkane preferably comprises dichloromethane and / or dichloroethane; the ester solvent preferably comprises ethyl acetate and / or butyl acetate. In the present invention, the volume ratio of the chloroalkane to the ester solvent in the mixed solvent is preferably 0.5 to 2:1, and in specific embodiments, it can be 0.5:1, 1:1, 1.5:1, or 2:1; when the mixed solvent of a chloroalkane and an ester solvent is used in the present invention, the yield of the target product is higher. In the present invention, the ratio of the amount of substance of Compound 1 to the volume of the organic solvent is preferably 1 mol:5 to 15 L, and in specific embodiments, it can be 1 mol:5 L, 1 mol:6 L, 1 mol:7 L, 1 mol:8 L, 1 mol:9 L, 1 mol:10 L, 1 mol:11 L, 1 mol:12 L, 1 mol:13 L, 1 mol:14 L, or 1 mol:15 L.

[0056] In the present invention, the temperature of the ring expansion reaction is preferably 80 to 150 °C, and in specific embodiments, it can be 80 °C, 90 °C, 100 °C, 110 °C, 120 °C, 130 °C, 140 °C or 150 °C; the time of the ring expansion reaction is preferably 10 to 20 h, and in specific embodiments, it can be 10 h, 11 h, 12 h, 13 h, 14 h, 15 h, 16 h, 17 h, 18 h, 19 h or 20 h. In the present invention, the ring expansion reaction is preferably carried out under a protective atmosphere, and the protective atmosphere preferably includes nitrogen, argon or helium. In the present invention, taking dichloromethane and ethyl acetate as organic solvents, the reaction formula of the ring expansion reaction is as follows:

[0057]

[0058] After completing the ring expansion reaction, the present invention preferably further includes: extracting the obtained ring expansion reaction solution with ethyl acetate and saturated brine, drying with anhydrous sodium sulfate, filtering to remove sodium sulfate, distilling off the solvent under reduced pressure, and performing silica gel column chromatography for separation and purification to obtain the benzonaphthyridin-5-one derivative. In the present invention, the eluent used for silica gel column chromatography separation and purification preferably includes a mixed solvent of petroleum ether and ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate in the mixed solvent is preferably 1 to 4:1, and in specific embodiments, it can be 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1 or 4:1.

[0059] The present invention also provides the application of the benzonaphthyridin-5-one derivative described in the above technical solution in the preparation of anti-tumor drugs, anti-inflammatory drugs, antibacterial drugs, antiviral drugs, neuroprotective drugs, drugs for reducing amyloid burden, drugs for treating cardiovascular diseases or organic electronic devices. In the present invention, the tumor preferably includes lung cancer.

[0060] In order to further illustrate the present invention, the following examples will be used to describe in detail a benzonaphthyridin-5-one derivative and its preparation method, application and pharmaceutical composition provided by the present invention, but they should not be construed as limiting the protection scope of the present invention.

[0061] Example 1

[0062]

[0063] Compound 1a (0.2 mmol), compound 2a (0.5 mmol), dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (0.004 mmol), copper(I) chloride (0.4 mmol), and solvents dichloroethane and ethyl acetate (volume ratio = 2:1, 2 mL) were added successively. The reaction was carried out under a nitrogen atmosphere at 120 °C with stirring for 10 h. The reaction mixture was extracted with ethyl acetate and saturated brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, the solvent was evaporated under reduced pressure, and the product was purified by silica gel column chromatography (petroleum ether:ethyl acetate volume ratio = 2:1) to obtain compound 3aa with a yield of 80%.

[0064] Figure 1 It is the 1H NMR spectrum of compound 3aa, Figure 2 and it is the 13C NMR spectrum of compound 3aa. 1 H NMR (400 MHz, CDCl3, ppm): δ 8.49 (s, 1H), 8.44 (d, J = 8.4 Hz, 1H), 8.24 - 8.22 (m, 2H), 7.69 - 7.66 (m, 1H), 7.61 - 7.59 (m, 2H), 7.55 - 7.45 (m, 6H), 7.43 - 7.37 (m, 2H), 3.69 (s, 3H); 13 C NMR (100 MHz, CDCl3, ppm): δ 163.4, 160.3, 157.2, 143.1, 142.7, 139.9, 138.5, 131.9, 129.8, 128.9, 128.6, 127.9, 127.6, 124.2, 122.5, 117.6, 116.9, 115.2, 110.3, 30.0. It can be seen that the target compound was successfully prepared in this invention.

[0065] Example 2

[0066]

[0067] Compound 1b (0.2 mmol), compound 2a (0.5 mmol), dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (0.004 mmol), copper(I) chloride (0.4 mmol), and solvents dichloroethane and ethyl acetate (volume ratio = 2:1, 2 mL) were added successively. The reaction was carried out under a nitrogen atmosphere at 120 °C with stirring for 10 h. The reaction mixture was extracted with ethyl acetate and saturated brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, the solvent was evaporated under reduced pressure, and the product was purified by silica gel column chromatography (petroleum ether:ethyl acetate volume ratio = 2:1) to obtain compound 3ba with a yield of 62%.

[0068] Figure 3 It is the 1H NMR spectrum of compound 3ba, Figure 4 and it is the 13C NMR spectrum of compound 3ba.1 HNMR (400 MHz, CDCl3, ppm): δ 9.54 (s, 1H), 8.22 (d, J = 6.8 Hz, 2H), 7.64 - 7.62 (m, 2H), 7.54 - 7.44 (m, 8H), 7.37 - 7.35 (m, 1H), 3.66 (s, 3H); 13 C NMR (100 MHz, CDCl3, ppm): δ 162.9, 159.8, 156.6, 143.2, 141.8, 141.7, 138.5, 133.3, 130.7, 129.9, 128.9, 128.9, 128.2, 127.6, 127.7, 126.5, 117.7, 116.1, 115.1, 114.0, 31.1. It can be seen that the target compound was successfully prepared in the present invention.

[0069] Example 3

[0070]

[0071] Compound 1c (0.2 mmol), compound 2a (0.5 mmol), dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (0.004 mmol), copper(I) chloride (0.4 mmol), solvent dichloroethane and ethyl acetate (volume ratio = 2:1, 2 mL) were added successively. The reaction was carried out under a nitrogen atmosphere at 120 °C with stirring for 10 h, extracted with ethyl acetate and saturated brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, the solvent was distilled off under reduced pressure, and purified by silica gel column chromatography (petroleum ether:ethyl acetate volume ratio = 2:1) to obtain compound 3ca with a yield of 70%.

[0072] Figure 5 is the 1H NMR spectrum of compound 3ca, Figure 6 is the 13C NMR spectrum of compound 3ca. 1 H NMR (400 MHz, CDCl3, ppm): δ 8.42 (s, 1H), 8.23 - 8.20 (m, 2H), 8.15 (s, 1H), 7.60 - 7.58 (m, 2H), 7.52 - 7.41 (m, 7H), 7.26 - 7.24 (m, 1H), 3.62 (s, 3H), 2.50 (s, 3H); 1313C NMR (100 MHz, CDCl3, ppm): δ 163.3, 160.1, 156.9, 143.2, 142.6, 138.6, 137.8, 132.9, 132.0, 129.8, 128.8, 128.7, 127.9, 127.6, 127.6, 124.2, 117.4, 117.1, 115.1, 110.2, 30.0, 20.9. It can be seen that the target compound was successfully prepared in the present invention.

[0073] Example 4

[0074]

[0075] Compound 1d (0.2 mmol), compound 2a (0.5 mmol), dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (0.004 mmol), copper(I) chloride (0.4 mmol), solvent dichloroethane and ethyl acetate (volume ratio = 2:1, 2 mL) were added successively. The reaction was carried out under a nitrogen atmosphere at 120 °C with stirring for 10 h, extracted with ethyl acetate and saturated brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, the solvent was distilled off under reduced pressure, and purified by silica gel column chromatography (petroleum ether:ethyl acetate volume ratio = 2:1) to obtain compound 3da with a yield of 70%.

[0076] Figure 7 is the 1H NMR spectrum of compound 3da, Figure 8 is the 13C NMR spectrum of compound 3da. 1 1H NMR (400 MHz, CDCl3, ppm): δ 8.54 (s, 1H), 8.47 (d, J = 8.0 Hz, 1H), 8.28 (d, J = 7.2 Hz, 2H), 7.68 - 7.67 (m, 2H), 7.59 - 7.46 (m, 7H), 7.37 - 7.22 (m, 7H), 5.59 - 5.58 (m, 2H); 13 13C NMR (100 MHz, CDCl3, ppm): δ 163.5, 160.5, 157.4, 143.1, 142.8, 139.2, 138.5, 136.4, 131.8, 129.9, 128.9, 128.9, 128.8, 128.1, 127.6, 127.2, 126.5, 124.2, 122.6, 117.9, 116.8, 116.2, 110.2, 46.2. It can be seen that the target compound was successfully prepared in the present invention.

[0077] Example 5

[0078]

[0079] Compound 1e (0.2 mmol), compound 2a (0.5 mmol), dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (0.004 mmol), copper(I) chloride (0.4 mmol), and the solvents dichloroethane and ethyl acetate (volume ratio = 2:1, 2 mL) were added successively. The reaction was carried out under a nitrogen atmosphere at 120 °C with stirring for 10 h. The reaction mixture was extracted with ethyl acetate and saturated brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, the solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate volume ratio = 2:1) to obtain compound 3ea in a yield of 70%.

[0080] Figure 9 This is the 1H NMR spectrum of compound 3ea, Figure 10 and this is the 13C NMR spectrum of compound 3ea. 1 H NMR (400 MHz, CDCl3, ppm): δ 8.46 (d, J = 6.8 Hz, 2H), 8.25 - 8.22 (m, 2H), 7.68 - 7.62 (m, 3H), 7.55 - 7.43 (m, 7H), 7.38 - 7.34 (m, 1H), 4.38 - 4.33 (m, 2H), 1.36 - 1.33 (m, 3H); 13 C NMR (100 MHz, CDCl3, ppm): δ 163.2, 159.8, 157.1, 143.1, 142.7, 138.8, 138.6, 131.9, 129.8, 128.9, 128.8, 128.0, 127.6, 124.5, 122.3, 117.9, 117.0, 115.1, 110.1, 37.7, 12.6. It can be seen that the target compound was successfully prepared in this invention.

[0081] Example 6

[0082]

[0083] Compound 1a (0.2 mmol), compound 2b (0.5 mmol), dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (0.004 mmol), copper(I) chloride (0.4 mmol), and the solvents dichloroethane and ethyl acetate (volume ratio = 2:1, 2 mL) were added successively. The reaction was carried out under a nitrogen atmosphere at 120 °C with stirring for 10 h. The reaction mixture was extracted with ethyl acetate and saturated brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, the solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate volume ratio = 2:1) to obtain compound 3ab in a yield of 76%.

[0084] Figure 11 This is the 1H NMR spectrum of compound 3ab, Figure 12 and this is the 13C NMR spectrum of compound 3ab.1 1H NMR (400 MHz, CDCl3, ppm): δ 8.40 - 8.35 (m, 2H), 8.20 (d, J = 8.8 Hz, 2H), 7.66 - 7.57 (m, 3H), 7.40 - 7.33 (m, 2H), 7.03 - 7.01 (m, 4H), 3.88 (d, J = 4.0 Hz, 6H), 3.69 (s, 3H); 13 13C NMR (100 MHz, CDCl3, ppm): δ 162.7, 161.2, 160.5, 159.6, 156.6, 142.7, 139.9, 135.5, 131.6, 131.1, 130.4, 129.1, 124.1, 122.4, 117.8, 116.1, 116.1, 114.2, 113.0, 108.7, 55.4, 55.3, 30.0. It can be seen that the target compound was successfully prepared in the present invention.

[0085] Example 7

[0086]

[0087] Compound 1a (0.2 mmol), compound 2c (0.5 mmol), dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (0.004 mmol), copper(I) chloride (0.4 mmol), and solvents dichloroethane and ethyl acetate (volume ratio = 2:1, 2 mL) were added successively. The reaction was carried out under a nitrogen atmosphere at 120 °C with stirring for 10 h, extracted with ethyl acetate and saturated brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the solvent was evaporated under reduced pressure. The product was purified by silica gel column chromatography (petroleum ether:ethyl acetate volume ratio = 2:1) to obtain compound 3ac with a yield of 64%.

[0088] Figure 13 It is the 1H NMR spectrum of compound 3ac, Figure 14 It is the 13C NMR spectrum of compound 3ac. 1 1H NMR (400 MHz, CDCl3, ppm): δ 8.51 (s, 1H), 8.38 (s, 1H), 8.23 (d, J = 7.6 Hz, 2H), 7.73 (d, J = 8.8 Hz, 1H), 7.59 - 7.47 (m, 8H), 7.29 - 7.26 (m, 1H), 3.66 (s, 3H); 1313C NMR (100 MHz, CDCl3, ppm): δ 163.5, 159.9, 157.6, 142.8, 141.5, 138.9, 138.2, 134.5, 130.0, 128.9, 128.6, 128.1, 127.6, 127.6, 126.9, 119.4, 116.9, 116.9, 115.6, 110.1, 77.4, 77.0, 76.7, 30.1. It can be seen that the target compound was successfully prepared in the present invention.

[0089] Example 8

[0090]

[0091] Compound 1a (0.2 mmol), compound 2d (0.5 mmol), dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (0.004 mmol), copper(I) chloride (0.4 mmol), solvent dichloroethane and ethyl acetate (volume ratio = 2:1, 2 mL) were added successively. The reaction was carried out under nitrogen atmosphere at 120 °C with stirring for 10 h, extracted with ethyl acetate and saturated brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, the solvent was evaporated under reduced pressure, and purified by silica gel column chromatography (petroleum ether:ethyl acetate volume ratio = 2:1) to obtain compound 3ad with a yield of 77%.

[0092] Figure 15 is the 1H NMR spectrum of compound 3ad, Figure 16 is the 13C NMR spectrum of compound 3ad. 1 1H NMR (400 MHz, CDCl3, ppm): δ 8.29 - 8.26 (m, 2H), 8.01 (d, J = 6.4 Hz, 2H), 7.53 - 7.49 (m, 1H), 7.41 - 7.39 (m, 2H), 7.27 - 7.14 (m, 6H), 3.56 (s, 3H), 3.33 (d, J = 14.4 Hz, 6H); 13 13C NMR (100 MHz, CDCl3, ppm): δ 163.2, 160.3, 157.04, 142.6, 140.3, 140.1, 140.0, 137.6, 135.7, 131.7, 129.547, 128.7, 128.4, 127.5, 124.2, 122.4, 117.8, 116.7, 115.1, 109.5, 30.0, 21.6, 21.4. It can be seen that the target compound was successfully prepared in the present invention.

[0093] Example 9

[0094]

[0095] Compound 1a (0.2 mmol), compound 2e (0.5 mmol), dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (0.004 mmol), copper(I) chloride (0.4 mmol), and a solvent mixture of dichloroethane and ethyl acetate (volume ratio = 2:1, 2 mL) were added successively. The reaction was carried out under a nitrogen atmosphere at 120 °C with stirring for 10 h. The reaction mixture was extracted with ethyl acetate and saturated brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate volume ratio = 2:1) to obtain compound 3ae with a yield of 66%.

[0096] Figure 17 This is the 1H NMR spectrum of compound 3ae, Figure 18 and this is the 13C NMR spectrum of compound 3ae. 1 HNMR (400 MHz, CDCl3, ppm): δ 8.35 - 8.33 (m, 2H), 7.93 - 7.85 (m, 2H), 7.58 - 7.54 (m, 1H), 7.32 - 7.26 (m, 3H), 7.21 - 7.14 (m, 3H), 3.60 (s, 3H), 2.28 - 2.25 (m, 12H); 13 C NMR (100 MHz, CDCl3, ppm): δ 163.4, 160.4, 157.3, 142.5, 141.0, 140.0, 138.6, 137.1, 136.3, 136.2, 135.8, 131.7, 130.1, 129.6, 128.9, 128.7, 126.2, 125.1, 124.2, 122.4, 117.8, 116.7, 115.1, 109.6, 77.4, 77.1, 76.7, 30.0, 20.1, 20.0, 19.9, 19.8. It can be seen that the target compound was successfully prepared in this invention.

[0097] Test Example 1

[0098] Human non-small cell lung cancer cell line (A549) was selected as the research object, and the CCK-8 method was used to preliminarily evaluate the anti-tumor activity of the synthesized benzonaphthyridin-5-one derivatives of this invention. Among them, A549 was purchased from Shanghai Huzhen Industrial Co., Ltd. The experimental steps are as follows:

[0099] 1. Cell resuscitation: Turn on the purification workbench and disinfect it with ultraviolet light for 30 min. Open the liquid nitrogen tank, find the cryopreservation tube of A549 cells. Take a beaker and add normal temperature water at 37 °C. Shake the cryopreservation tube left and right in the beaker to quickly thaw it. Then dry the surface and transfer it to a centrifuge. Centrifuge at 1000 r / min for 4 min. Take it out and transfer it to the purification workbench. First, disinfect the outer surface of the cryopreservation tube with medical alcohol. Open it, discard the upper layer of cryopreservation liquid, add 1 mL of culture medium again, gently and slowly disperse the cells with a pipette, and transfer them to a 25 cm 2 culture flask. Then pipette 4 mL of culture medium and add it to the culture flask. Observe under an inverted microscope and find that the appearance and morphology of the cells are normal. Transfer them to a constant temperature incubator with 5% carbon dioxide at 37 °C. After 24 h, observe the cells, aspirate the culture medium, and replace it with fresh culture medium. In this operation step, the bottle mouth and bottle cap of the culture flask need to be sterilized by passing through the outer flame of an alcohol lamp when opening and closing.

[0100] 2. Cell passage: Observe the cells under a microscope and find that they have basically covered the bottom surface of the culture flask and can be passaged. Before the operation, irradiate the purification workbench and the consumables used in the experiment with ultraviolet light for 30 min for disinfection. Open the culture flask and aspirate the culture solution with a pipette. Pipette 3 mL of buffer solution to wash the bottom cells twice, aspirate it, add 1 mL of 0.25% trypsin, and gently shake the bottle body horizontally. Observe the cells again and find that they show a tendency of cytoplasmic retraction. Immediately stop digestion, add 2 mL of RPMI1640 culture medium, slowly blow down the adherent cells, transfer the cell solution to a centrifuge, centrifuge at 1000 r / min for 4 min, aspirate the upper layer of culture medium, add 1.5 mL of culture medium again, slowly and gently disperse the cells, divide the cell solution into 3 culture flasks, add 5 mL of culture medium to the culture flasks again, observe the cells in the culture flasks under a microscope, and there is no abnormal situation. Disinfect the outer surface of the bottle body with medical alcohol and transfer it to a constant temperature incubator with 5% carbon dioxide at 37 °C.

[0101] 3. Cell cryopreservation: Observe under a microscope and select cells in the logarithmic growth phase. Aspirate the upper layer of culture solution, add 1 mL of 0.25% trypsin, observe that the cells show a tendency of cytoplasmic retraction, and terminate digestion. Add 1.5 mL of culture medium, gently blow down the adherent cells, transfer them to a centrifuge, centrifuge at 1000 r / min for 3 min, aspirate the culture medium, add an appropriate amount of cryopreservation solution, and divide and transfer it into sterile cryopreservation tubes, about 1.5 mL per tube. The processed cryopreservation tubes are first temporarily stored in a 4 °C refrigerator for 40 min, transferred to a -20 °C refrigerator for 2.5 h, then to an -80 °C refrigerator for 12 h, and finally transferred to the liquid nitrogen tank.

[0102] 4. Cell viability detection: Observe under a microscope that human lung cancer cells A549 basically cover more than 80% of the bottom of the culture flask and are in good condition. Aspirate the culture medium, wash it twice with PBS buffer solution, digest it with trypsin for 1 min, then add an appropriate amount of culture medium to dilute the cells, and perform counting to make the cell density about 4×103 cells / mL. After adding 100 μL of buffer to the periphery of a 96-well plate, add 100 μL of cell suspension to each of the remaining wells, and transfer it to an incubator for culture. Add 100 μL of medium to the blank group, and add 100 μL of medium containing different monomer benzonaphthyridin-5-one derivatives at concentrations (0.2 μmol / L, 0.4 μmol / L, 0.6 μmol / L, 0.8 μmol / L, 1 μmol / L, 2 μmol / L, 3 μmol / L, 4 μmol / L) to the experimental group. Set 3 replicate wells, culture for 48 h, add 10 μL of CCK-8 indicator to each well, culture for 4 h, then transfer it to an enzyme-linked immunosorbent assay (ELISA) reader for detection. Set the wavelength to 450 nm, and process the data according to the measured absorbance OD value. By performing different concentration gradient treatments on each compound, the corresponding half-maximal inhibitory concentration (IC 50 ) value was calculated to evaluate its inhibitory effect on tumor cell proliferation.

[0103] 5. Experimental results: Use Graphpad prism software to process the experimental data of the compounds, fit the trend change line graph of the dependence relationship between the experimental concentration and the inhibition rate, and then the IC 50 value corresponding to each monomer compound can be obtained. The results are shown in Table 1.

[0104] Table 1 IC 50 values of benzonaphthyridin-5-one derivatives

[0105] Compound <![CDATA[A549IC 50 (μM)]]> Compound <![CDATA[A549IC 50 (μM)]]> 3aa 83.25±7.04 3ab 49.84±4.73 3ca 122.16±8.41 3ad 93.25±5.14 3da 108.19±5.66 3ae 87.25±6.23 3ea 103.25±6.03

[0106] As can be seen from Table 1, among the benzonaphthyridin-5-one derivatives provided by the present invention, compound 3ab (methoxy-substituted benzonaphthyridin-5-one derivative) showed the most significant inhibitory effect, with the lowest IC 50 value, indicating strong anti-tumor potential; the IC 50 values of the remaining compounds were relatively low, showing a certain inhibitory effect, indicating that these compounds have moderate cytotoxicity.

[0107] The above preliminary results indicate that benzonaphthyridin-5-one derivatives have potential pharmacological activities in inhibiting the proliferation of non-small cell lung cancer cells in vitro. In particular, the activity of compound 3ab is worthy of further in-depth study. It provides an important basis for subsequent chemical structure optimization and the development of more potent anti-tumor candidate compounds, and also lays a foundation for clarifying the mechanism of action of these compounds and their application potential in clinical practice.

[0108] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A benzonaphthyridin-5-one derivative, characterized in that, It has the structure shown in Formula I: Among them, R 1 is hydrogen, methyl, methoxy or chlorine; R 2 is benzyl, methyl or ethyl; R 3 is hydrogen, methyl, methoxy or chlorine, and the number of R 3 on a single benzene ring is 1 to 2.

2. The benzonaphthyridin-5-one derivative according to claim 1, wherein The benzonaphthyridin-5-one derivative has the structure shown in Formula 3aa, Formula 3ba, Formula 3ca, Formula 3da, Formula 3ea, Formula 3ab, Formula 3ac, Formula 3ad or Formula 3ae:

3. The method for preparing the benzonaphthyridin-5-one derivative according to claim 1 or 2, comprising the following steps: Subjecting Compound 1 and Compound 2 to a ring-expansion reaction in the presence of an organometallic catalyst to obtain the benzonaphthyridin-5-one derivative; The organometallic catalyst is dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer; the ring-expansion reaction is carried out in the presence of an additive, and the additive is copper(I) chloride; The organic solvent used in the ring-expansion reaction is dichloroethane and ethyl acetate.

4. The preparation method according to claim 3, wherein The molar ratio of Compound 1 to the organometallic catalyst is 1:0.01 - 0.

05.

5. The preparation method according to claim 3, characterized in that The molar ratio of Compound 1 to the additive is 1:0.5 - 2.

5.

6. The preparation method according to claim 3, wherein The molar ratio of Compound 1 to Compound 2 is 1:1.25 - 5.

7. The preparation method according to any one of claims 3 to 6, characterized in that, The temperature of the ring-expansion reaction is 80 - 150 °C, and the time is 10 - 20 h; The ring-expansion reaction is carried out under a protective atmosphere.

8. Use of the benzonaphthyridin-5-one derivative according to any one of claims 1 - 2 in the preparation of an anti-tumor drug.

Images