Spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives and their synthesis methods and anticancer applications

Abstract

The present invention provides a class of spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3'-indole derivatives and their synthesis methods and anticancer applications. 2-hydroxy-1,4-naphthoquinone, isatin derivatives and aminopyrazole compounds are selected as raw materials, β-1-methylimidazole-2,3,4,6-tetrasulfonic acid-D-pyranose glucose sulfhydrate is used as a catalyst, and the reaction is obtained under mild conditions. Two new C-N bonds, four new C-C bonds, a new dihydropyridine ring and a spiro ring structure are formed in one step. The reaction conditions are mild, the substrate range is wide, the yield is high, and the post-processing is convenient. All raw materials are commercially available. The obtained product has a complex structure and multiple biologically active sites. At 12.5 μg / mL, the inhibition rate of 7 compounds on MCF-7 cells is higher than 30% (2 compounds are greater than or equal to 90%); the inhibition rate of 10 compounds on MHCC-97H cells is higher than 30%.

Description

Technical Field

[0001] The present invention belongs to the field of chemical synthesis, and specifically relates to spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives, a synthesis method thereof and anti-tumor activity thereof. Background Art

[0002] Compounds with a spirocyclic skeleton are subunits that make up many alkaloids, are templates for new drug development, and are used as scaffolds for combinatorial libraries.

[0003] Indole and dihydroindole are important components of natural products and drugs. Connecting dihydroindole, spirocyclic ring, etc. to the 3rd carbon of the indole ring can show efficient biological activity. Due to the presence of the indole part, spiro-oxindole compounds have various biological activities and are the core structure of many drugs and natural alkaloids.

[0004] Multicomponent reaction is a classic method for synthesizing spirooxindole compounds, and its core element is the catalyst. The reported catalysts include weak acids such as proline, p-toluenesulfonic acid, medelnic acid, and cyanuric chloride. Probably due to the insufficient acidity of these catalysts, the corresponding reactions must be carried out under high temperature conditions and only moderate yields can be obtained; and these reactions have very strict requirements on the electronic effect and steric hindrance of the substrate. Substrates with large steric hindrance cannot react smoothly, resulting in a narrow substrate range and the structural diversity of the products needs to be broadened. Therefore, it is of great significance to develop novel, general, and efficient methods for the synthesis of spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives.

[0005] Sugar sulfonic acid is a compound synthesized from natural glucose. Since there are four hydroxyl groups on the sugar ring, the derived sugar sulfonic acid has four sulfonic acid groups in its molecular structure, and its acidity is equivalent to that of inorganic strong acid. And its unique chair structure allows the four sulfonic acid groups to perform their respective functions while acting synergistically, so it can play the role of bringing different raw materials together, greatly enhancing the effective collision probability of raw material molecules, allowing the reaction to proceed smoothly under milder conditions, while improving the yield and broadening the substrate range, obtaining products with complex structures, novel structures and good diversity.

[0006] In view of the structural advantages of sugar sulfonic acid, the present invention uses sugar sulfonic acid β-1-methylimidazole-2,3,4,6-tetrasulfonic acid-D-pyranose glucose sulfhydrate as a catalyst, 2-hydroxy-1,4-naphthoquinone, isatin derivatives and aminopyrazole compounds as raw materials, and reacts under mild conditions to obtain spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives. Summary of the invention

[0007] The purpose of the present invention is to synthesize spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives by using 2-hydroxy-1,4-naphthoquinone, isatin derivatives and aminopyrazole compounds as raw materials and β-1-methylimidazole-2,3,4,6-tetrasulfonic acid-D-pyranose glucose hydrogen sulfide as a catalyst through a one-step reaction.

[0008] A class of spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives, having the general structural formula:

[0009]

[0010] Among them, R 1 OCH 3 , -H, -F, -Cl, -NO 2 、-CH 3 Any one of R 2 -CH 3 , Any of R 3 -H or -CH 3 .

[0011] The spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives described above have a structural formula of any one of the following:

[0012]

[0013]

[0014] The synthesis method of the spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives comprises the following steps:

[0015]

[0016] After the reaction was completed, the mixture was cooled to room temperature and filtered. The product was washed with water and ethanol to obtain a red solid crude product, which was purified by column chromatography to obtain product 4.

[0017] The reagent used in column chromatography is V 石油醚 :V 乙酸乙酯 =4:1.

[0018] The invention relates to the use of the spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives in the preparation of anticancer drugs.

[0019] The application of the spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives in the preparation of drugs for treating liver cancer and breast cancer.

[0020] The application described herein is directed to the inhibition of proliferation of human liver cancer MHCC-97H cells, and the application described herein is directed to the inhibition of proliferation of human breast cancer triple-positive MCF-7 cells.

[0021] Beneficial effects:

[0022] 1. Using β-1-methylimidazole-2,3,4,6-tetrasulfonic acid-D-pyranose glucose sulfhydrate as catalyst, 2-hydroxy-1,4-naphthoquinone, isatin derivatives and aminopyrazole compounds as raw materials, spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives were constructed in the next step under mild conditions.

[0023] 2. The synthesis method of the present invention forms two new CN bonds, four new CC bonds, a new dihydropyridine ring and a spiro ring structure in one step.

[0024] 3. The synthesis method of the present invention has a wide range of substrates (indigo carmine derivatives with electron-withdrawing and electron-donating groups, and aminopyrazole derivatives with large steric substituents at the 1,3-positions can all react) and a high yield.

[0025] 4. All raw materials in the synthesis method of the present invention are commercially available and do not need to be synthesized in advance. The reaction conditions are mild (50°C) and the post-treatment is convenient, which provides an efficient and rapid route for the synthesis of spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives.

[0026] 5. The obtained products have a complex structure, with a tetracyclic fused and spirocyclic structure, and have multiple biologically active sites. At 12.5 μg / mL, the inhibition rate of 7 compounds on MCF-7 cells was higher than 30% (2 compounds were greater than or equal to 90%); the inhibition rate of 10 compounds on MHCC-97H cells was higher than 30%, so they have excellent anti-tumor activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The H NMR spectrum of the catalyst (400 MHz, DMSO-d 6 );

[0028] Figure 2 The carbon NMR spectrum of the catalyst (100 MHz, DMSO-d 6 );

[0029] Figure 3 The H NMR spectrum of product 4a (400 MHz, DMSO-d 6 );

[0030] Figure 4 The carbon NMR spectrum of product 4a (100 MHz, DMSO-d6 );

[0031] Figure 5 This is the HRMS (ESI) spectrum of product 4a. DETAILED DESCRIPTION

[0032] The present invention will be further described below in conjunction with the embodiments.

[0033] Embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention. It will be appreciated by those skilled in the art that various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention.

[0034] A class of spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivatives, whose structural formula is one of the following:

[0035]

[0036] The present invention also provides a method for preparing the novel spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivative, comprising:

[0037]

[0038] 2-Hydroxy-1,4-naphthoquinone 1 (1 mmol), isatin derivative 2 (1 mmol), aminopyrazole 3 (1 mmol), catalyst β-1-methylimidazole-2,3,4,6-tetrasulfonic acid-D-pyranose glucose sulfhydrate (15 mol%, homemade, synthesis method see authorized patent ZL201510103426.4) and ethanol (3 mL) were added into a 10 mL round-bottom flask in sequence and reacted at 50 °C until completion (TLC tracking monitoring, according to the R f The product was cooled to room temperature, filtered, and washed with a small amount of water and ethanol to obtain a red crude product. 石油醚 :V 乙酸乙酯 =4:1) to obtain product 4.

[0039] Table 1 Synthesis of spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivative 4

[0040]

[0041] The application of the spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole derivative in the preparation of anticancer drugs.

[0042] Example 1 Synthesis of Compound 5′-methoxy-3-methyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione (4a):

[0043] 2-Hydroxy-1,4-naphthoquinone (1mmol), 5-methoxyisatin (1mmol), 3-methyl-1-phenyl-1H-pyrazole-5-amine (1mmol), catalyst β-1-methylimidazole-2,3,4,6-tetrasulfonic acid-D-pyranose glucose hydrogensulfide (15mol%) and ethanol (3mL) were added to a 10mL round-bottom flask in sequence, and reacted at 50°C until complete (TLC tracking monitoring). Cooled to room temperature, filtered, the product was washed with a small amount of water and ethanol to obtain a red powdery solid crude product, which was purified by column chromatography (V 石油醚 :V 乙酸乙酯 =4:1) to obtain pure product 4a. The specific reaction route is as follows:

[0044]

[0045] Example 2 Synthesis of Compound 3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione (4b):

[0046] 2-Hydroxy-1,4-naphthoquinone (1mmol), isatin (1mmol), 1-phenyl-3-(p-tolyl)-1H-pyrazole-5-amine (1mmol), catalyst β-1-methylimidazole-2,3,4,6-tetrasulfonic acid-D-pyranose glucose hydrogensulfide (15mol%) and ethanol (3mL) were added to a 10mL round-bottom flask in sequence, and reacted at 50°C until complete (TLC tracking monitoring). Cooled to room temperature, filtered, the product was washed with a small amount of water and ethanol to obtain a red powdery solid crude product, which was purified by column chromatography (V 石油醚 :V 乙酸乙酯 =4:1) to obtain pure product 4b. The specific reaction route is as follows:

[0047]

[0048] Example 3 Synthesis of Compound 3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione (4h):

[0049] 2-Hydroxy-1,4-naphthoquinone (1mmol), isatin (1mmol), 3-(4-methoxyphenyl)-1-phenyl-1H-pyrazole-5-amine (1mmol), catalyst β-1-methylimidazole-2,3,4,6-tetrasulfonic acid-D-pyranose glucose hydrogensulfide (15mol%) and ethanol (3mL) were added to a 10mL round-bottom flask in sequence, and reacted at 50°C until complete (TLC tracking monitoring). Cooled to room temperature, filtered, the product was washed with a small amount of water and ethanol to obtain a red powdery solid crude product, which was purified by column chromatography (V 石油醚 :V 乙酸乙酯 =4:1) to obtain pure product 4h. The specific reaction route is as follows:

[0050]

[0051] Example 4 Synthesis of Compound 3-methyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione (4n):

[0052] 2-Hydroxy-1,4-naphthoquinone (1mmol), isatin (1mmol), 3-methyl-1-(p-tolyl)-1H-pyrazole-5-amine (1mmol), catalyst β-1-methylimidazole-2,3,4,6-tetrasulfonic acid-D-pyranose glucose hydrogensulfide (15mol%) and ethanol (3mL) were added to a 10mL round-bottom flask in sequence, and reacted at 50°C until complete (TLC tracking monitoring). Cooled to room temperature, filtered, the product was washed with a small amount of water and ethanol to obtain a red powdery solid crude product, which was purified by column chromatography (V 石油醚 :V 乙酸乙酯 =4:1) to obtain pure product 4n. The specific reaction scheme is as follows:

[0053]

[0054] The synthesis methods of the remaining products are similar to those of 4a, 4b, 4h and 4n. 5-methoxyisatin is replaced with isatin and its derivatives in the general formula;

[0055] The properties of the following compounds are described in detail below:

[0056]

[0057] 1. Compound 4a

[0058] The molecular formula is: C 29 H 20 N 4 O 4

[0059] Chinese name: 5′-methoxy-3-methyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0060] English name: 5′-Methoxy-3-methyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0061] Appearance: red solid

[0062] Melting point: 240.3-242.1℃

[0063] Infrared spectrum: IR(KBr)(v max ,cm -1 )3426,3343,2938,2360,2342,2209,1755,1727,1678.

[0064] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ1.65(3H,s,CH 3 ),3.61(3H,s,OCH 3 ),6.69-8.08(12H,m,Ar-H),9.78(1H,s,NH),10.51(1H,s,NH).

[0065] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.6,179.4,178.7,155.1,145.0,141.0,138.2,138.1,137.9,137.8,136.0,135.2,134.7,133.2, 131.8,129.9,129.5,127.2,126.0,125.8,122.9,115.2,112.5,111.1,109.3,101.3,55.3,50.3,11.3.

[0066] 2. Compound 4b

[0067] The molecular formula is: C 34 H 22 N 4 O 3

[0068] Chinese name: 3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0069] English name:

[0070] 3-p-Methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0071] Appearance: red solid

[0072] Melting point: 243.1-244.6℃

[0073] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3151,3082,2940,2202,1710,1640,1593.

[0074] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ2.27(3H,s,CH 3 ),6.57-8.07(17H,m,Ar-H),9.79(1H,s,NH),10.17(1H,s,NH).

[0075] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.4,179.4,178.7,149.3,142.0,140.6,138.1,137.9,137.2,136.4,135.2,133.3,131.8,129.7,129 .6,129.5,128.4,128.2,127.9,127.6,125.9,125.9,124.0,123.1,121.5,115.5,109.2,101.1,50.0,20.8.

[0076] 3. Compound 4c

[0077] The molecular formula is: C 34 H 21 N 4 O 3 F

[0078] Chinese name: 5′-fluoro-3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0079] English name:

[0080] 5′-Fluoro-3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-

[0081] 2′,5,10(1H,11H)-trione

[0082] Appearance: red solid

[0083] Melting point: 254.2-256.1℃

[0084] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3592,3342,2951,2212,1711,1679,1649.

[0085] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ2.28(3H,s,CH 3 ),6.55-8.08(16H,m,Ar-H),9.93(1H,s,NH),10.20(1H,s,NH).

[0086] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.7,179.3,178.8,159.4,157.1,144.9,141.2,138.1,137.6,136.2,135.2,133.3,131.7,129.9,129.5,127.3, 126.0,125.8,123.0,119.9,118.6,116.2,116.1,114.6,114.4,114.2,112.0,111.7,109.7,109.6,100.8,50.3,11.3.

[0087] 4. Compound 4d

[0088] The molecular formula is: C 34 H 21 N 4 O 3Cl

[0089] Chinese name: 5′-chloro-3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0090] English name:

[0091] 5′-Chloro-3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0092] Appearance: red solid

[0093] Melting point: 261.9-263.7℃

[0094] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3564,3394,2931,2212,1715,1674,1646.

[0095] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ2.29(3H,s,CH 3 ),6.93-8.08(16H,m,Ar-H),9.93(1H,s,NH),10.31(1H,s,NH).

[0096] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.4,179.4,178.7,149.3,142.0,140.6,138.1,137.9,137.2,136.4,135.2,133.3,131.8,129.7, 129.6,129.5,128.4,128.2,127.9,127.6,125.9,124.0,123.1,121.5,115.5,109.2,101.1,50.0,20.8.

[0097] 5. Compound 4e

[0098] The molecular formula is: C 34 H 21 N 5 O 5

[0099] Chinese name: 5′-nitro-3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0100] English name:

[0101] 5′-Nitro-3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0102] Appearance: red solid

[0103] Melting point: 239.2-241.1℃

[0104] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3422,3191,2947,2207,1734,1680,1620,1536,1352.

[0105] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ2.29(3H,s,CH 3 ),6.62-8.12(16H,m,Ar-H),10.08(1H,s,NH),10.88(1H,s,NH).

[0106] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.4,179.8,179.2,159.4,149.7,142.4,141.1,138.5,138.3,136.7,135.7,133.8,132.3,130.2,130 .2,130.1,128.7,128.1,126.4,126.4,125.0,124.5,123.6,122.0,116.0,113.3,109.7,101.7,55.5,20.7.

[0107] 6. Compound 4f

[0108] The molecular formula is: C 35 H 24 N 4 O 3

[0109] Chinese name: 5′-methyl-3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0110] English name:

[0111] 5′-Methyl-3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0112] Appearance: red solid

[0113] Melting point: 241.6-243.3℃

[0114] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3420,3062,2985,2361,1755,1727,1621.

[0115] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ2.19(3H,s,CH 3 ),2.28(3H,s,CH 3 ),6.54-8.06(16H,m,Ar-H),9.83(1H,s,NH),10.07(1H,s,NH).

[0116] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.4,179.4,178.6,155.0,149.5,140.5,139.0,137.9,137.5,136.3,135.7,133.7,132.3,129 .8,129.3,129.2,129.1,127.9,127.0,125.4,124.1,122.6,115.2,112.3,108.5,100.8,55.5,20.0.

[0117] 7. Compound 4g

[0118] The molecular formula is: C 35 H 24 N 4 O 4

[0119] Chinese name: 5′-methoxy-3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0120] English name:

[0121] 5′-Methoxy-3-p-methylphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0122] Appearance: red solid

[0123] Melting point: 212.3-214.2℃

[0124] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3426,3343,2921,2842,2363,2218,1755,1727,1678.

[0125] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ2.28(3H,s,CH 3 ),3.64(3H,s,OCH 3 ),6.54-8.08(12H,m,Ar-H),9.81(1H,s,NH),10.00(1H,s,NH).

[0126] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.4,179.4,178.6,155.0,149.4,140.5,139.0,138.0,137.2,136.3,135.4,135.2,133.2,131.8,129 .7,129.6,129.4,128.4,127.9,127.6,125.9,123.1,115.5,112.8,111.2,109.5,101.0,55.4,50.5,20.8.

[0127] 8. Compound 4h

[0128] The molecular formula is: C 34 H 22 N 4 O4

[0129] Chinese name: 3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0130] English name:

[0131] 3-p-Methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0132] Appearance: red solid

[0133] Melting point: 222.8-224.6℃

[0134] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3151,3082,2936,2210,1716,1660,1607.

[0135] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ3.74(3H,s,OCH 3 ),6.58-8.07(17H,m,Ar-H),9.86(1H,s,NH),10.16(1H,s,NH).

[0136] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.0,178.8,178.7,158.9,149.1,142.0,138.2,138.2,138.1,137.9,136.2,135.2,131.9,129.7,129 .5,128.6,128.3,127.5,125.9,125.8,124.0,123.1,112.8,109.2,109.2,102.6,101.3,100.2,55.0,50.0.

[0137] 9. Compound 4i

[0138] The molecular formula is: C 34 H 21 N 4 O 4 F

[0139] Chinese name: 5′-fluoro-3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0140] English name:

[0141] 5′-Fluoro-3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0142] Appearance: red solid

[0143] Melting point: 240.5-242.0℃

[0144] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3592,3442,2996,2215,1711,1679,1642.

[0145] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ3.75(3H,s,OCH 3 ),6.55-8.08(16H,m,Ar-H),9.93(1H,s,NH),10.18(1H,s,NH).

[0146] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ181.0,179.8,179.3,159.7,159.5,149.7,141.3,139.8,139.5,138.6,138.6,136.7,135.7,133 .8,132.3,130.2,130.1,128.1,126.4,124.9,123.7,115.4,113.3,112.1,110.2,101.4,55.5,51.0.

[0147] 10. Compound 4j

[0148] The molecular formula is: C 34 H 21 N 4 O 4 Cl

[0149] Chinese name: 5′-chloro-3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0150] English name:

[0151] 5′-Chloro-3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0152] Appearance: red solid

[0153] Melting point: 237.4-238.6℃

[0154] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3564,3394,2985,2209,1715,1674,1645.

[0155] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ3.75(3H,s,OCH 3 ),6.60-8.08(16H,m,Ar-H),9.91(1H,s,NH),10.29(1H,s,NH).

[0156] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ181.0,179.8,179.0,159.5,149.6,141.4,139.9,138.5,136.8,135.7,133.8,132.3,130.3,130 .2,130.0,128.4,128.1,126.4,125.8,124.9,124.6,123.7,115.2,113.4,111.0,101.3,55.5,50.7.

[0157] 11. Compound 4k

[0158] The molecular formula is: C 34 H 21 N 5 O 6

[0159] Chinese name: 5′-nitro-3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0160] English name:

[0161] 5′-Nitro-3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0162] Appearance: red solid

[0163] Melting point: 217.3-219.2℃

[0164] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3422,3191,2990,2212,1734,1680,1620,1558,1354.

[0165] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ3.73(3H,s,CH 3 ),6.57-8.07(16H,m,Ar-H),9.87(1H,s,NH),10.16(1H,s,NH).

[0166] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.9,179.8,179.2,159.4,149.7,142.4,141.1,138.5,138.3,136.7,135.7,133.8,132.3,130.2,130 .2,130.1,128.7,128.1,126.4,126.4,125.0,124.5,123.6,122.0,116.0,113.3,109.7,101.7,55.5,50.5.

[0167] 12. Compound 4l

[0168] The molecular formula is: C 35 H 24 N 4 O 4

[0169] Chinese name: 5′-methyl-3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0170] English name:

[0171] 5′-Methyl-3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0172] Appearance: red solid

[0173] Melting point: 254.3-256.3℃

[0174] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3420,3062,3002,2987,2360,2209,1755,1727,1621.

[0175] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ2.19(3H,s,CH 3 ),3.74(3H,s,OCH 3 ),6.55-8.07(16H,m,Ar-H),9.83(1H,s,NH),10.05(1H,s,NH).

[0176] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.9,179.9,179.1,159.4,149.6,141.0,140.0,138.6,138.5,136.7,135.7,133.7,132.3,130.8, 130.3,130.2,130.1,128.9,128.0,126.4,125.1,123.6,116.2,113.3,109.5,101.8,55.5,50.6,21.0.

[0177] 13. Compound 4m

[0178] The molecular formula is: C 35 H 24 N 4 O 5

[0179] Chinese name: 5′-methoxy-3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0180] English name:

[0181] 5′-Methoxy-3-p-methoxyphenyl-1-phenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0182] Appearance: red solid

[0183] Melting point: 216.4-218.1℃

[0184] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3426,3343,2998,2945,2836,2202,1755,1727,1678.

[0185] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ1.65(3H,s,CH 3 ),3.61(3H,s,OCH 3 ),6.69-8.08(16H,m,Ar-H),9.78(1H,s,NH),10.51(1H,s,NH).

[0186] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.9,179.9,179.0,159.4,155.4,149.7,141.1,139.4,138.6,136.7,135.9,135.7,133.7,132.4, 130.3,130.2,130.1,128.0,126.4,125.1,123.6,116.0,113.2,111.6,110.0,101.7,55.9,55.5,51.0.

[0187] 14. Compound 4n

[0188] The molecular formula is: C 29 H 20 N 4O 3

[0189] Chinese name: 3-methyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0190] English name:

[0191] 3-Methyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0192] Appearance: red solid

[0193] Melting point: 244.8-247.6℃

[0194] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3151,3082,2921,2859,2363,2218,1710,1675,1632.

[0195] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ2.40(3H,s,CH 3 ),3.36(3H,s,CH 3 ),6.86-8.06(12H,m,Ar-H),9.69(1H,s,NH),10.67(1H,s,NH).

[0196] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.6,179.3,178.8,144.7,141.3,140.9,136.8,136.7,135.9,135.6,135.2,133.3,131.7, 129.9,129.8,128.1,126.0,125.8,123.9,122.9,121.8,115.2,109.0,101.1,49.8,20.5,11.2.

[0197] 15. Compound 4o

[0198] The molecular formula is: C 29 H 19 N 4 O 3F

[0199] Chinese name: 5′-fluoro-3-methyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0200] English name:

[0201] 5′-Fluoro-3-methyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0202] Appearance: red solid

[0203] Melting point: 198.7-200.7℃

[0204] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3592,3442,2947,2361,2343,2207,1711,1679,1642.

[0205] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ1.64(3H,s,CH 3 ),2.40(3H,s,CH 3 ),6.89-8.07(11H,m,Ar-H),9.74(1H,s,NH),10.70(1H,s,NH).

[0206] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.7,179.3,178.9,144.6,141.1,138.2,138.1,137.5,137.0,136.0,135.6,135.2,133.3,131.8, 129.9,129.8,128.6,126.0,125.8,123.0,114.5,114.4,114.2,109.7,109.7,100.6,50.3,20.5,11.2.

[0207] 16. Compound 4p

[0208] The molecular formula is: C 29 H 19 N 4O 3 Cl

[0209] Chinese name: 5′-chloro-3-methyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0210] English name:

[0211] 5′-Chloro-3-methyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0212] Appearance: red solid

[0213] Melting point: 252.4-254.2℃

[0214] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3564,3394,2934,2361,2212,1715,1674,1645.

[0215] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ1.65(3H,s,CH 3 ),2.40(3H,s,CH 3 ),6.92-8.07(11H,m,Ar-H),9.74(1H,s,NH),10.81(1H,s,NH).

[0216] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.7,179.3,178.9,144.6,141.1,138.2,138.1,137.5,137.0,136.0,135.6,135.2,133.3,131.8, 129.9,129.9,128.6,126.0,125.9,123.0,114.6,114.4,114.2,109.7,109.7,100.6,50.3,20.6,11.2.

[0217] 17. Compound 4q

[0218] The molecular formula is: C 29 H 19 N5 O 5

[0219] Chinese name: 5′-nitro-3-methyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0220] English name:

[0221] 5′-Nitro-3-methyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0222] Appearance: red solid

[0223] Melting point: 238.3-240.1℃

[0224] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3422,3191,2996,2211,1734,1680,1620,1545,1352.

[0225] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ1.65(3H,s,CH 3 ),2.41(3H,s,CH 3 ),7.14-8.21(11H,m,Ar-H),9.91(1H,s,NH),11.44(1H,s,NH).

[0226] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.8,179.4,179.1,147.8,144.5,142.5,141.5,137.2,137.1,136.2,135.5,135.2,133.4 ,131.6,130.0,129.9,126.1,125.9,125.7,123.2,119.8,113.8,109.2,99.9,49.8,20.6,11.3.

[0227] 18. Compound 4r

[0228] The molecular formula is: C 30 H 22 N4 O 3

[0229] Chinese name: 3,5′-dimethyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0230] English name:

[0231] 3,5′-Dimethyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0232] Appearance: red solid

[0233] Melting point: 207.9-209.6℃

[0234] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3420,3062,2990,2885,2336,2212,2107,1755,1727,1621.

[0235] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ1.63(3H,s,CH 3 ),2.14(3H,s,CH 3 ),2.40(3H,s,CH 3 ),6.79-8.06(11H,m,Ar-H),9.68(1H,s,NH),10.57(1H,s,NH).

[0236] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.6,179.4,178.9,144.7,140.9,138.8,136.8,136.8,135.9,135.6,135.2,133.3,131.7,13 0.7,129.9,129.8,128.3,126.0,125.8,124.5,122.9,115.3,108.8,101.2,49.8,20.5,20.5,11.2.

[0237] 19. Compound 4s

[0238] The molecular formula is: C 30 H 22 N 4 O 4

[0239] Chinese name: 5′-methoxy-3-methyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indole]-2′,5,10(1H,11H)-trione

[0240] English name:

[0241] 5′-Methoxy-3-methyl-1-p-methylphenylspiro[benzo[h]pyrazolo[3,4-b]quinoline-4,3′-indoline]-2′,5,10(1H,11H)-trione

[0242] Appearance: red solid

[0243] Melting point: 240.3-242.1℃

[0244] Infrared spectrum: IR(KBr)(v max ,cm -1 ):3426,3343,3219,2991,2836,2202,1755,1727,1678.

[0245] H NMR spectrum: 1 H NMR (400 MHz, DMSO-d 6 ):δ1.64(3H,s,CH 3 ),2.40(3H,s,CH 3 ),3.60(3H,s,OCH 3 ),6.68-8.07(11H,m,Ar-H),9.65(1H,s,NH),10.49(1H,s,NH).

[0246] C NMR spectrum: 13 C NMR (100 MHz, DMSO-d 6 ): δ180.6,179.4,178.7,155.1,144.7,140.9,137.9,136.8,135.9,135.7,135.2,134.6,133.2,13 1.8,129.9,129.8,126.0,125.8,122.9,115.1,112.5,111.1,109.3,101.1,55.3,50.3,20.5,11.3.

[0247] The physical properties (yield, melting point, etc.) of each compound are shown in Table 1.

[0248] Example 2 Antitumor Activity Assay

[0249] In this experiment, the Alamar Blue method was used to test the antitumor activity of product 4 using cell lines (human normal liver LO2 cell line, human breast cancer triple-positive MCF-7 cell line, human breast cancer triple-negative MDA-MB-231 cell line, human liver cancer HepG2 cell line and human liver cancer MHCC-97H cell line). The inhibition rates of different concentrations of compound 4 on each cell line are as follows (Tables 2-6).

[0250] Table 2 Effects of different concentrations of compound 4 on the proliferation of normal human liver LO2 cells a

[0251]

[0252] a Inhibition rate less than 30% is marked as -

[0253] From the results in Table 2, it can be seen that the inhibition rate of 12.5 μg / mL of compounds 4c-4o on the proliferation of normal human liver LO2 cells is low, especially 4f has no inhibition on normal cells at a concentration of up to 50 μg / mL, indicating that its toxicity is very low.

[0254] Table 3 Effects of different concentrations of compound 4 on the proliferation of triple-positive human breast cancer MCF-7 cells a

[0255]

[0256]

[0257] a Inhibition rate less than 30% is marked as -

[0258] From the results in Table 3, it can be seen that 12.5 μg / mL of 4n-4p and 4r-4s have a significant improvement in the inhibition rate of proliferation of triple-positive human breast cancer MCF-7 cells, especially 4r-4s, whose inhibition rate on triple-positive human breast cancer MCF-7 cells is greater than or equal to 90%, which is of great significance for further research.

[0259] Table 4 Effects of different concentrations of compound 12 on the proliferation of triple-negative breast cancer MDA-MB-231 cells a

[0260]

[0261] a Inhibition rates below 30% are marked as –

[0262] The results in Table 4 indicate that this type of compound is not a suitable inhibitor of triple-negative human breast cancer MDA-MB-231 cells.

[0263] Table 5 Effects of different concentrations of compound 4 on the proliferation of human liver cancer HepG2 cells a

[0264]

[0265] a Inhibition rate less than 30% is marked as -

[0266] From the results in Table 5, it can be seen that this type of compound has no inhibitory effect on the proliferation of human liver cancer HepG2 cells.

[0267] Table 6 Effects of different concentrations of compound 4 on the proliferation of human hepatocellular carcinoma MHCC-97H cells a

[0268]

[0269]

[0270] a Inhibition rate less than 30% is marked as -

[0271] From the results in Table 6, it can be seen that the inhibition rates of the 10 compounds on the proliferation of human liver cancer MHCC-97H cells at 12.5 μg / mL were higher than those on normal human liver LO2 cells, especially the inhibition rates of 4r and 4s exceeded 75%, which is worthy of further study.

Claims

1. A spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3'-indole derivative, characterized in that: The general structural formula is: Wherein, R1 is any one of -OCH3, -H, -F, -Cl, -NO2, -CH3; R2 is any one of -CH3, Any one of; R3 is -H or -CH3; but when R2 is -CH3, R1 and R3 are not -H at the same time.

2. A spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3'-indole derivative according to claim 1, characterized in that: The structural formula is any one of the following:

3. A method for synthesizing a spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3'-indole derivative according to claim 1 or 2, characterized in that: The following steps are involved: The amount of cat. used is 15 mol%.

4. The method for synthesizing a spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3'-indole derivative according to claim 3, characterized in that: After the reaction was completed, the mixture was cooled to room temperature and filtered. The product was washed with water and ethanol to obtain a red solid crude product, which was purified by column chromatography to obtain product 4.

5. The method for synthesizing a spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3'-indole derivative according to claim 3, characterized in that: The reagent used in column chromatography is V 石油醚 :V 乙酸乙酯 =4:

1.

6. Use of the spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3'-indole derivatives according to claim 1 or 2 in the preparation of anticancer drugs.

7. Use of the spirobenzo[h]pyrazolo[3,4-b]quinoline-4,3'-indole derivatives according to claim 1 or 2 in the preparation of drugs for treating liver cancer and breast cancer.

8. The use according to claim 7, characterized in that: The liver cancer is aimed at inhibiting the proliferation of human liver cancer MHCC-97H cells, and the breast cancer is aimed at inhibiting the proliferation of human breast cancer triple-positive MCF-7 cells.

Images