CN114057755B

CN114057755B - Preparation method and application of iridoid aglycone-based spiro indolone compound

Info

Publication number: CN114057755B
Application number: CN202111206613.7A
Authority: CN
Inventors: 穆淑珍; 何茂飞; 范艳华; 邓璐璐; 徐兴连; 李江; 郝小江
Original assignee: Key Laboratory of Natural Product Chemistry of Guizhou Academy of Sciences
Current assignee: Key Laboratory of Natural Product Chemistry of Guizhou Academy of Sciences
Priority date: 2021-10-17
Filing date: 2021-10-17
Publication date: 2022-08-30
Anticipated expiration: 2041-10-17
Also published as: CN114057755A

Abstract

The invention discloses a preparation method and application of a spiro indolone compound based on natural iridoid aglycone, belonging to the field of drug synthesis. The invention synthesizes the spiro indolone compound by natural iridoid aglycone, diethyl aminomalonate and benzylidene indolone under the catalysis of BINOLs phosphoric acid for the first time, and the structure of the spiro indolone compound is shown as a formula I and a formula II. The preparation method has the characteristics of simplicity and easy operation, can be realized through one-step or several-step limited reaction, and does not have high temperatureHigh pressure and extreme reaction of strong acid and strong alkali, simple and easily available reaction raw materials, and in vitro tumor activity test and mechanism research on the reaction raw materials. The compound of the invention has stronger inhibiting effect on tumor cells and can be used as a lead compound of antitumor drugs.

Description

Preparation method and application of iridoid aglycone-based spiro indolone compound

Technical Field

The invention relates to the field of organic chemistry and pharmaceutical chemistry, in particular to a preparation method and application of a spiro-indolone compound based on iridoid aglycone.

Background

Cancer is a common and frequently occurring disease that is severely life threatening to humans, with mortality second to cardiovascular disease. Due to population growth and aging, the incidence of cancer increases by 33% between 2005 and 2015. According to the international cancer research institute report, about 960 ten thousand cancer deaths in 2018, it is expected that about 2150 ten thousand new cancer cases will occur in 2025. Cancer is becoming the first killer of humans and one of the biggest public health problems worldwide. Today, chinese cancer presents a trend of three high rates of youthfulness, morbidity, and mortality. Therefore, research and development of high-efficiency anticancer drugs have become a problem to be solved urgently.

Genipin is a hydrolysis product of geniposide, and is mainly present in Chinese traditional medicinal plants such as gardenia, eucommia and the like, and both have various pharmacological activities. This limits the application of genipin aglycone in the medical field. Therefore, if the genipin aglycone is subjected to reasonable structural modification design and synthesis, some chemical entities with novel structures and higher antitumor activity are synthesized, and the method has important research significance for discovering innovative drug candidate molecules.

Spirocyclic indolone alkaloids exhibit varying degrees of anticancer activity, which activity is attributed primarily to the spiro substitution at the C3 position of the oxoindole and to the substituents on the oxoindole. Many of the natural spirocyclic indolone compounds, such as spirotropic statin sA and B, also exhibit good anticancer activity. More importantly, some compounds with spiro indolone as core structure have entered preclinical research.

Disclosure of Invention

In order to solve the problem of the deficiency of the prior art, the invention aims to provide a preparation method and application of a spiro-indolone alkaloid based on iridoid aglycone. The compounds have obvious inhibition effect on tumor cells.

In order to achieve the purpose, the invention adopts the technical scheme that: iridoid aglycone-based spiro indolone compounds have structures shown in formula I and formula II:

wherein R is selected from one of hydrogen, halogen and trifluoromethyl; r ₁ Selected from hydrogen, halogen, trifluoromethyl, methoxy, carboxy, trifluoromethoxy; r is ₂ Selected from methyl, morpholine ring, tetrahydropyrrole ring; r ₃ Selected from hydrogen, acetyl; x is selected from N or CH.

Wherein R is selected from halogen, methoxy, trifluoromethyl and trifluoromethoxy.

The compounds of formula I and formula II are selected from the following compounds:

the invention discloses a preparation method of a compound shown in formula (I), which comprises the following steps of reacting a compound shown in formula (III-1) with compounds shown in formula (a) and formula (b) to generate a compound shown in formula (I):

wherein R is selected from H, chlorine and trifluoromethyl; r is ₁ Selected from hydrogen, halogen, trifluoromethyl, methoxy, carboxy, trifluoromethoxy; r ₂ Selected from methyl, morpholine rings, tetrahydropyrrole rings; r ₃ Selected from hydrogen, acetyl; x is selected from N or CH.

The invention relates to a preparation method of a compound shown in a formula (II), which comprises the following steps of reacting a compound shown in a formula (III-2) with compounds shown in formulas (c) and (b) to generate a compound shown in the formula (II):

The beneficial technical effects of the invention are as follows: the invention firstly takes genipin aglycone as a synthesis block, and synthesizes the spiro indolone compound with genipin aglycone through 1, 3-dipolar cycloaddition reaction under the catalysis of phosphoric acid derived from BINOLs. The in vitro test results show that the compounds have stronger inhibitory activity on human non-small cell lung cancer HCC827, human highly metastatic liver cancer cell MHCC97H and human neuroblastoma SHSY-5Y.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without inventive step based on the embodiments of the present invention, are within the scope of protection of the present invention.

Example 1

The procedure for the preparation of methyl (1R, 4aS, 7aS) -7-formyl-1-methoxy-1, 4a,5,7 a-tetrahydrocyclopent [ c ] pyran-4-carboxylate, described in this example:

genipin (Genipin) (1g,4.5mmol) was weighed into a 25mL round bottom flask, 4mL methanol was added, a drop of concentrated HCl was added, and the reaction was stirred at 60 ℃ for 6 h. Adding 1mol/L sodium hydroxide into the reaction solution to adjust the pH value to 7, concentrating under reduced pressure, extracting the residue with ethyl acetate, washing the extract with saturated saline solution, drying with anhydrous sodium sulfate, and evaporating the solvent under reduced pressure. 20mL of dried methylene chloride was added to dissolve the resulting solution, and 3g (34mmol) of activated manganese dioxide was added to the solution to react at room temperature for 24 hours. Manganese dioxide was removed by filtration, the solvent was removed under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate: 8:1, V/V) to give two isomers, the configuration of which was confirmed by 2D-NOESY, respectively, in which the target product a was the main product, 250mg of a white solid, yield 60%, ¹ H NMR(600MHz,CDCl ₃ )δ2.44-2.54(m,1H),3.04-3.14(m,1H),3.25(dt,J＝2.7,5.6Hz,1H),3.29(s,3H),3.76(s,3H),5.28(d,J＝3.8Hz,1H),7.10-7.03(m,1H),7.47(d,J＝1.3Hz,1H),9.77(s,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ32.66,40.52,42.15,51.26,56.29,98.14,110.45,145.29,150.70,155.77,167.71,189.33.MS(ESI)m/z:261.3[M+Na] ⁺ 。

example 2

The preparation method of (1R, 4aS, 7aS) -1-methoxy-4- (morpholine-4-carbonyl) -1,4a,5,7 a-tetrahydro-chloropentano [ c ] pyran-7-carbaldehyde described in this example:

compound M1(1g, 4.2mmol) was dissolved in 5M KOH ethanol solution, refluxed for 2h, concentrated under reduced pressure, redissolved in ethyl acetate, washed with 1N hydrochloric acid solution, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give compound 3; add Compound C (200mg, 0.88mmol), HoBt (338mg, 3.16mmol), EDCI (480mg,3.16mmol) and TEA (40mg, 0.40mmol) to dichloromethane at 0 deg.C; the mixture was stirred for 10min, then morpholine (280mg, 3.17mmol) was added to the solution and stirred at room temperature overnight; adding water into the reaction solution, and extracting with dichloromethane for three times; washing the organic layer with saturated brine, drying over anhydrous sodium sulfate, and concentrating under reduced pressure to obtain compound D; dissolving the compound D in dichloromethane, adding activated MnO ₂ (3.6g, 42mmol), stirring at room temperature for 24 h; filtering the mixture, vacuum concentrating, and purifying with silica gel column chromatography (petroleum ether: ethyl acetate: 2:1, V/V) to obtain Ea as main product; 500mg of a colorless oily liquid, yield 46%, MS (ESI) M/z 294.0[ M + H ]] ⁺ 。

Example 3

The preparation method of (1R, 4aS, 7aS) -1-methoxy-4- (pyrrolidine-1-carbonyl) -1,4a,5,7 a-tetrahydro-chloropenta [ c ] pyran-7-carbaldehyde described in this example:

compound M1(1g, 4.2mmol) is dissolved in 5M KOH ethanol solution, refluxed for 2h, concentrated under reduced pressure, redissolved in ethyl acetate, washed with 1N hydrochloric acid solution and saturatedWashed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give compound 3. Compound C (200mg, 0.88mmol) was added to dichloromethane at 0 deg.C, and HoBt (338mg, 3.16mmol), EDCI (480mg,3.16mmol) and TEA (40mg, 0.40mmol) were added. The mixture was stirred for 10min, then pyrrolidine (225mg, 3.17mmol) was added to the solution and stirred at room temperature overnight; water was added to the reaction solution, and the mixture was extracted three times with methylene chloride. Washing the organic layer with saturated brine, drying over anhydrous sodium sulfate, and concentrating under reduced pressure to obtain compound D; dissolving the compound D in dichloromethane, adding activated MnO ₂ (3.6g, 42mmol), stirring at room temperature for 24 h; the mixture was filtered, concentrated in vacuo and purified by silica gel column chromatography (petroleum ether: ethyl acetate: 2:1, V/V) to give Ga as the major product. 500mg of a colorless oily liquid, yield 50%, MS (ESI) M/z 278.0[ M + H ]] ⁺ 。

Example 4

The preparation of (E) -1-acetyl-3-benzylideneindol-2-one described in this example:

a round-bottomed flask was charged with 2-indolone (396mg,3.0mmol), benzaldehyde (350mg,3.3mmol) and 4mL of anhydrous ethanol, and piperidine (52 mg, 0.6mmol) was slowly added dropwise and heated at 80 ℃ for 2 hours. Precipitate is generated, and the precipitate is filtered and washed by ethanol to obtain (E) -3-benzylidene indole-2-ketone. (E) -3-benzylideneindol-2-one was dissolved in 5mL of tetrahydrofuran, acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol) were added, stirred at room temperature for 12h, diluted with water, and extracted with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2X 100mL) and then anhydrous MgSO ₄ The crude product was dried and purified on a silica gel column (petroleum ether: ethyl acetate 60:1, V/V) to give the title compound (542 mg of yellow solid, 54% yield). And (3) product identification result: m.p.145-146 ℃; ¹ H NMR(600MHz,CDCl ₃ )δ8.34(d,J＝8.2Hz,1H),7.91(s,1H),7.72(d,J＝7.5Hz,1H),7.66(d,J＝7.1Hz,2H),7.50(ddd,JJ＝8.5,7.6,6.2Hz,3H),7.37–7.32(m,1H),7.05(td,J＝7.7,0.9Hz,1H),2.79(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ170.91,168.62,140.28,138.71,134.42,130.28,130.07,129.21,128.81,124.49,122.16,121.88,116.75,27.00,26.93；HRMS(ESI):Calcd.for C ₁₇ H ₁₃ O ₂ NNa[M+Na] ⁺ :286.0838；found:286.0831。

example 5

The procedure for the preparation of (E) -1-acetyl-6-chloro-3- (3 (trifluoromethyl) benzylidene) indol-2-one described in this example:

preparation method As in example 4, 6-chloro-2-indolone (500mg,3.0mmol), 3-trifluoromethylbenzaldehyde (574mg,3.3mmol) and 4mL of anhydrous ethanol were added to a round-bottomed flask, and 52mg of piperidine (0.6mmol) was slowly added dropwise and heated at 80 ℃ for 2 hours. Precipitate is generated, and the precipitate is filtered and washed by ethanol to obtain (E) -6-chloro-3- (3 (trifluoromethyl) benzylidene) indol-2-one. (E) -6-chloro-3- (3 (trifluoromethyl) benzylidene) indol-2-one was dissolved in 5mL tetrahydrofuran, acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol) were added, stirred at room temperature for 12h, diluted with water and extracted with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2X 100mL) and then anhydrous MgSO ₄ Drying and purification of the crude product on silica gel column (petroleum ether: ethyl acetate 60:1, V/V) gave the title compound (321 mg of yellow solid, 31% yield). And (3) product identification result: m.p.142-143 ℃; ¹ H NMR(600MHz,CDCl ₃ )δ8.35(dd,J＝11.7,4.6Hz,3H),7.73(d,J＝8.1Hz,1H),7.62(t,J＝7.9Hz,1H),7.60(s,1H),7.53(d,J＝8.2Hz,1H),7.25(dd,J＝8.2,1.9Hz,1H),2.74(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ170.84,165.69,139.53,136.87,135.84,134.59,133.67,128.91,128.81,128.49,128.46,127.32,127.30,125.61,125.16,122.65,119.78,117.32,26.86；HRMS(ESI):Calcd.for C ₁₈ H ₁₁ O ₂ NClF ₃ Na[M+Na] ⁺ :388.0323；found:388.0311。

example 6

The preparation of (E) -1-acetyl-5-chloro-3- (3-chloro-2-fluorobenzylidene) indol-2-one described in this example:

preparation was as in example 4, 5-chloro-2-indolone (500mg,3.0mmol), 2-fluoro-3-chlorobenzaldehyde (521mg,3.3mmol) and 4mL of absolute ethanol were added to a round-bottomed flask, and 52mg (0.6mmol) of piperidine was slowly added dropwise and heated at 80 ℃ for 2 hours. Precipitate is generated, and the precipitate is filtered and washed by ethanol to obtain (E) -5-chloro-3- (2-fluoro-3-chlorobenzylidene) indol-2-one. Dissolve (E) -5-chloro-3- (2-fluoro-3-chlorobenzylidene) indol-2-one in 5mL tetrahydrofuran, add acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol), stir at room temperature for 12h, add water for dilution, and extract with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2X 100mL), and then with anhydrous MgSO ₄ Drying and purification of the crude product on silica gel (petroleum ether: ethyl acetate 60:1, V/V) afforded the title compound (268 mg of yellow solid, 27% yield). And (3) product identification result: m.p.141-142 ℃; ¹ H NMR(600MHz,CDCl ₃ )δ8.30(d,J＝8.8Hz,1H),7.87(s,1H),7.59–7.55(m,2H),7.43(d,J＝1.9Hz,1H),7.36–7.31(m,1H),7.26(t,J＝7.9Hz,1H),2.78(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ170.55,167.23,139.09,132.80,131.16,131.15,130.69,130.22,130.16,130.13,128.04,127.68,124.88,124.85,122.70,122.40,119.55,118.07,26.79；HRMS(ESI):Calcd.for C ₁₈ H ₁₀ O ₂ NC _l2 FNa[M+Na] ⁺ :372.0073；found:372.0083。

example 7

The procedure for the preparation of (E) -1-acetyl-6-chloro-3- (4- (trifluoromethyl) benzylidene) indol-2-one described in this example:

preparation method As in example 4, 6-chloro-2-indolone (500mg,3.0mmol), 4-trifluoromethylbenzaldehyde (574mg,3.3mmol) and 4mL of absolute ethanol were added to a round-bottomed flask, piperidine (52 mg, 0.6mmol) was slowly added dropwise, and 80 ℃ was addedHeating for 2 h. Precipitate is generated, and the precipitate is filtered and washed by ethanol to obtain (E) -6-chloro-3- (4- (trifluoromethyl) benzylidene) indol-2-one. (E) -6-chloro-3- (4- (trifluoromethyl) benzylidene) indol-2-one was dissolved in 5mL tetrahydrofuran, acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol) were added, stirred at room temperature for 12h, diluted with water, and extracted with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2X 100mL) and then anhydrous MgSO ₄ Drying and purification of the crude product on silica gel column (petroleum ether: ethyl acetate 60:1, V/V) gave the title compound (255 mg of yellow solid, 25% yield). And (3) identifying a product: m.p.143-145 ℃; ¹ H NMR(600MHz,CDCl ₃ )δ8.43(d,J＝1.9Hz,1H),7.84(s,1H),7.59–7.51(m,3H),7.49(s,1H),7.35(d,J＝6.7Hz,1H),7.06(dd,J＝8.4,2.0Hz,1H),2.78(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ168.22,159.84,140.92,139.07,135.88,135.38,130.05,125.32,124.64,123.10,121.49,117.28,116.10,114.23,77.25,77.04,76.83,55.42,26.82；HRMS(ESI):Calcd.for C ₁₈ H ₁₁ O ₂ NClF ₃ Na[M+Na] ⁺ :388.0323；found:388.0311。

example 8

The procedure for the preparation of (E) -1-acetyl-6-chloro-3- (3- (methoxy) benzylidene) indol-2-one described in this example:

preparation method 6-chloro-2-indolone (500mg,3.0mmol), 3-methoxybenzaldehyde (449mg,3.3mmol) and 4mL of absolute ethanol were added to a round-bottomed flask, 52mg (0.6mmol) of piperidine was slowly added dropwise, and the mixture was heated at 80 ℃ for 2 hours. Precipitate is generated, and the precipitate is filtered and washed by ethanol to obtain (E) -6-chloro-3- (3- (methoxyl) benzylidene) indole-2-ketone. (E) -6-chloro-3- (3- (methoxy) benzylidene) indol-2-one was dissolved in 5mL tetrahydrofuran, acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol) were added, stirred at room temperature for 12h, diluted with water and extracted with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2X 100mL) and then anhydrous MgSO ₄ Drying to obtain crude productPurification on a gel column (petroleum ether: ethyl acetate 60:1, V/V) gave the title compound (311 mg of yellow solid, 32% yield). And (3) product identification result: m.p.141-143 ℃; ¹ H NMR(600MHz,CDCl ₃ )δ8.39(d,J＝1.9Hz,1H),7.88(s,1H),7.67(d,J＝8.4Hz,1H),7.42(t,J＝7.9Hz,1H),7.21(d,J＝7.5Hz,1H),7.13(s,1H),7.06–6.99(m,2H),3.86(s,3H),2.77(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ168.22,159.84,140.92,139.07,135.88,135.38,130.05,125.32,124.64,123.10,121.49,117.28,116.10,114.23,77.25,77.04,76.83,55.42,26.82；HRMS(ESI):Calcd.for C ₁₈ H ₁₄ O ₃ NClNa[M+Na] ⁺ :350.0554；found:350.0546。

example 9

The preparation method of the (E) -1-acetyl-6-chloro-3- (pyridine-4-yl methylene) indole-2 ketone comprises the following steps:

preparation method As in example 4, 6-chloro-2-indolone (500mg,3.0mmol), 4-pyridinecarboxaldehyde (353mg,3.3mmol) and 4mL of absolute ethanol were added to a round-bottomed flask, 52mg of piperidine (0.6mmol) was slowly added dropwise, and the mixture was heated at 80 ℃ for 2 hours. Precipitate is generated, and the precipitate is filtered and washed by ethanol to obtain (E) -6-chloro-3- (pyridine-4-yl methylene) indole-2-ketone. (E) -6-chloro-3- (pyridin-4-ylmethylene) indol-2-one was dissolved in 5mL tetrahydrofuran, acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol) were added, stirred at room temperature for 12h, diluted with water, and extracted with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2X 100mL) and then anhydrous MgSO ₄ Drying and purification of the crude product on silica gel column (petroleum ether: ethyl acetate 60:1, V/V) gave the title compound (295 mg of yellow solid, 33% yield). And (3) product identification result: m.p.138-139 ℃; ¹ H NMR(600MHz,CDCl ₃ )δ8.79(d,J＝5.8Hz,2H),8.39(s,1H),7.76(s,1H),7.46(d,J＝5.5Hz,1H),7.40(d,J＝8.4Hz,1H),7.04(dd,J＝8.4,1.9Hz,2H),2.77(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ170.91,168.62,140.28,138.71,134.42,130.28,130.07,129.21,128.81,124.49,122.16,121.88,116.75,27.00,26.93；HRMS(ESI):Calcd.for C ₁₆ H ₁₂ O ₂ N ₂ Cl[M+H] ⁺ :299.0582；found:299.0576。

example 10

The preparation method of the (E) -1-acetyl-6-chloro-3- (3-methoxy-5- (trifluoromethyl) benzylidene) indole-2-ketone comprises the following steps:

preparation was as in example 4, 6-chloro-2-indolone (500mg,3.0mmol), 3-methoxy-5-trifluoromethylbenzaldehyde (673mg,3.3mmol) and 4mL of absolute ethanol were added in a round-bottomed flask, and 52mg of piperidine (0.6mmol) was slowly added dropwise and heated at 80 ℃ for 2 h. Precipitate is generated, and the precipitate is filtered and washed by ethanol to obtain (E) -6-chloro-3- (3-methoxy-5- (trifluoromethyl) benzylidene) indol-2-one. (E) -6-chloro-3- (3-methoxy-5- (trifluoromethyl) benzylidene) indol-2-one was dissolved in 5mL of tetrahydrofuran, acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol) were added, stirred at room temperature for 12h, diluted with water, and extracted with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2X 100mL) and then anhydrous MgSO ₄ Drying and purification of the crude product on silica gel column (petroleum ether: ethyl acetate 60:1, V/V) gave the title compound (296 mg of yellow solid, 25% yield). And (3) product identification result: m.p.140-142 ℃; ¹ H NMR(600MHz,CDCl ₃ )δ8.40(d,J＝1.9Hz,1H),7.83(s,1H),7.52–7.49(m,1H),7.23(s,2H),7.06(dd,J＝8.4,2.0Hz,1H),3.91(s,4H),2.77(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ170.86,170.59,167.82,165.68,160.16,159.66,141.27,137.02,136.58,136.26,126.71,124.84,122.99,119.74,119.70,119.61,117.91,117.88,117.59,117.52,117.27,112.26,112.23,55.80,26.79；HRMS(ESI):Calcd.for C ₁₉ H ₁₄ O ₃ NClF ₃ [M+H] ⁺ :396.0609；found:396.0616。

example 11

The procedure for the preparation of (E) -1-acetyl-6-chloro-3- (3- (trifluoromethoxy) benzylidene) indol-2-one described in this example:

preparation method As in example 4, 6-chloro-2-indolone (500mg,3.0mmol), 3-trifluoromethoxybenzaldehyde (627mg,3.3mmol) and 4mL of anhydrous ethanol were added to a round-bottomed flask, 52mg of piperidine (0.6mmol) was slowly added dropwise, and the mixture was heated at 80 ℃ for 2 hours. Precipitate is generated, and the precipitate is filtered and washed by ethanol to obtain (E) -6-chloro-3- (3- (trifluoromethoxy) benzylidene) indol-2-one. (E) -6-chloro-3- (3- (trifluoromethoxy) benzylidene) indol-2-one was dissolved in 5mL tetrahydrofuran, acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol) were added, stirred at room temperature for 12h, diluted with water, and extracted with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2 × 100mL), then dried over anhydrous MgSO4, and the crude product was purified on a silica gel column (petroleum ether: ethyl acetate 60:1, V/V) to give the title compound (302 mg of yellow solid, 26% yield). And (3) identifying a product: m.p.135-136 ℃; ¹ H NMR(600MHz,CDCl ₃ )δ8.41(d,J＝1.9Hz,1H),7.84(s,1H),7.59–7.51(m,3H),7.49(s,1H),7.35(d,J＝6.7Hz,1H),7.06(dd,J＝8.4,2.0Hz,1H),2.78(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ170.61,167.89,141.22,136.58,136.53,136.12,130.60,127.48,126.50,124.81,122.94,122.53,121.28,119.74,117.49,26.87,26.80；HRMS(ESI):Calcd.for C ₁₈ H ₁₂ O ₃ NClF ₃ [M+H] ⁺ :382.0452；found:382.03441。

example 12

The preparation of tert-butyl (E) -4- ((1-acetyl-6-chloro-2-oxoindole-3-propylidene) methyl) benzoate described in this example was carried out:

preparation method As in example 4, 6-chloro-2-indolone (500mg,3.0mmol), tert-butyl 4-formylbenzoate (680mg,3.3mmol) and 4mL of absolute ethanol were added to a round-bottomed flask, 52mg of piperidine (0.6mmol) was slowly added dropwise,heating at 80 deg.C for 2 h. Precipitate is generated, and the precipitate is filtered and washed by ethanol to obtain tert-butyl (E) -4- ((6-chloro-2-oxyindole-3-propylene) methyl) benzoate. Tert-butyl (E) -4- ((6-chloro-2-oxoindol-3-propylidene) methyl) benzoate was dissolved in 5mL of tetrahydrofuran, acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol) were added, stirred at room temperature for 12h, diluted with water, and extracted with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2 × 100mL), then dried over anhydrous MgSO4, and the crude product was purified on a silica gel column (petroleum ether: ethyl acetate 60:1, V/V) to give the title compound (368 mg of yellow solid, 31% yield). And (3) product identification result: m.p.138-139 ℃; ¹ H NMR(600MHz,CDCl ₃ )δ8.39(d,J＝1.9Hz,1H),8.11(d,J＝8.3Hz,2H),7.89(s,1H),7.66(d,J＝8.1Hz,2H),7.53(d,J＝8.4Hz,1H),7.03(dd,J＝8.4,2.0Hz,1H),2.77(s,3H),1.65(s,9H)； ¹³ C NMR(151MHz,CDCl ₃ )δ170.63,167.94,164.91,141.15,138.12,137.60,136.37,133.32,129.93,128.87,126.40,124.78,123.04,119.95,117.41,81.72,77.25,77.04,76.83,28.19,26.80；HRMS(ESI):Calcd.for C ₂₂ H ₂₀ O ₄ NClNa[M+Na] ⁺ :420.1081；found:420.1073。

example 13

The procedure for the preparation of (E) -1-acetyl-3- (3- (trifluoromethyl) benzylidene) indol-2-one described in this example:

preparation method As in example 4, 2-indolone (396mg,3.0mmol), 3-trifluoromethylbenzaldehyde (574mg,3.3mmol) and 4mL of absolute ethanol were added to a round-bottomed flask, and 52mg of piperidine (0.6mmol) was slowly added dropwise and heated at 80 ℃ for 2 hours. Precipitate is generated, and the precipitate is filtered and washed by ethanol to obtain (E) -3- (3- (trifluoromethyl) benzylidene) indol-2-one. (E) -3- (3- (trifluoromethyl) benzylidene) indol-2-one was dissolved in 5mL tetrahydrofuran, acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol) were added, stirred at room temperature for 12h, diluted with water and extracted with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2X 100mL) and then withDry over anhydrous MgSO4 and purify the crude product on silica gel column (petroleum ether: ethyl acetate 60:1, V/V) to give the title compound (284 mg of yellow solid, 29% yield). And (3) product identification result: m.p.121-123 ℃; ¹ H NMR(600MHz,CDCl ₃ )δ8.36(d,J＝8.2Hz,1H),7.92(s,1H),7.87(s,1H),7.84(d,J＝7.7Hz,1H),7.74(d,J＝7.8Hz,1H),7.65(dd,J＝16.0,8.4Hz,1H),7.56(d,J＝7.8Hz,1H),7.38(t,J＝7.6Hz,1H),7.07(t,J＝7.6Hz,1H),2.80(s,3H)； ¹³ C NMR(151MHz,CDCl ₃ )δ170.78,168.21,140.66,136.05,135.27,132.19,130.93,129.45,127.61,126.44,126.41,125.90,125.88,124.68,122.08,121.28,117.00,116.81,26.98,26.92；HRMS(ESI):Calcd.For C ₁₈ H ₁₂ O ₂ NF ₃ Na[M+Na] ⁺ :354.0712；found:354.0705。

example 14

The procedure for the preparation of (E) -1-acetyl-3- (3-chloro-2-fluorobenzylidene) -6- (trifluoromethyl) indol-2-one described in this example:

preparation method As in example 4, 6-trifluoromethyl-2-indolone (603mg,3.0mmol), 2-fluoro-3-chlorobenzaldehyde (521mg,3.3mmol) and 4mL of absolute ethanol were added to a round-bottomed flask, 52mg (0.6mmol) of piperidine was slowly added dropwise, and the mixture was heated at 80 ℃ for 2 hours. Precipitate is generated, and the precipitate is filtered and washed by ethanol to obtain (E) -3- (3-chloro-2-fluorobenzylidene) -6- (trifluoromethyl) indol-2-one. (E) -3- (3-chloro-2-fluorobenzylidene) -6- (trifluoromethyl) indol-2-one was dissolved in 5mL of tetrahydrofuran, acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol) were added, the mixture was stirred at room temperature for 12h, diluted with water, and extracted with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2X 100mL), and then with anhydrous MgSO ₄ Drying and purification of the crude product on silica gel column (petroleum ether: ethyl acetate 60:1, V/V) gave the title compound (384 mg as a yellow solid, 32% yield). And (3) product identification result: m.p.120-122 ℃; ¹ H NMR(600MHz,CDCl ₃ )δ2.81(s,3H),7.25(t,J＝7.8Hz,1H),7.36(dd,J＝0.7,8.1Hz,1H),7.56(d,J＝7.9Hz,2H),7.58(d,J＝8.4Hz,1H),7.95(s,1H),8.66(s,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ26.80,114.04,119.60,121.70,122.66,123.52,124.25,124.90,127.7,128.16,130.26,130.97,132.38,132.88,133.41,140.73,156.1,167.13,170.72.HRMS(ESI):Calcd.For C ₁₈ H ₁₀ O ₂ NClF ₄ Na[M+Na] ⁺ :429.0710；found:429.0705。

example 15

The procedure for the preparation of methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) -1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] pyran-4-carboxylate, described in this example:

6-chloro-2-indolone (500mg,3mmol), Compound A (785mg,3.3mmol) and 4mL of methanol were added to a round-bottomed flask, piperidine 52mg (0.6mmol) was slowly added dropwise, and stirring was carried out at room temperature for 24 hours, whereupon a precipitate was formed, which was filtered and washed with methanol to give methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) -1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] c]Pyran-4-carboxylic acid esters. This (1.2g,3.0mmol) was dissolved in 5mL tetrahydrofuran, acetic anhydride (4.5mL,15.0mmol) and sodium carbonate (156mg,15.0mmol) were added, stirred at room temperature for 12h, diluted with water, and extracted with ethyl acetate (3X 50 mL). The organic layer was washed with saturated brine (2 × 100mL), then dried over anhydrous sodium sulfate, and the crude product was purified on a silica gel column (petroleum ether: ethyl acetate ═ 10:1, V/V) to give the title compound (380 mg of yellow solid, 30%). And (3) product identification result: m.p.136-137 deg.c; ¹ H NMR(600MHz,CDCl ₃ )δ8.38(d,J＝1.9Hz,1H),7.78(d,J＝8.3Hz,1H),7.56(s,1H),7.42(s,1H),7.12(dd,J＝8.3,1.9Hz,1H),6.57(s,1H),4.65(d,J＝7.0Hz,1H),3.78(s,3H),3.54(s,3H),3.37(q,J＝7.8Hz,1H),3.17(dd,J＝18.5,8.4Hz,1H),2.97(t,J＝7.3Hz,1H),2.74(s,3H),2.47–2.40(m,1H)； ¹³ C NMR(151MHz,CDCl ₃ )δ170.74,168.06,167.55,152.33,140.70,138.79,138.07,135.53,134.78,125.29,124.70,122.99,120.73,117.29,110.56,101.90,57.25,51.39,49.14,40.43,34.48,26.78；HRMS(ESI):Calcd.for C ₂₂ H ₂₀ O ₆ NClNa[M+Na] ⁺ :452.0871；found:452.0858。

example 16

The preparation method of BN according to the embodiment comprises the following steps:

the method comprises the following steps: r-binaphthol (5g,1.8mmol) and 160mL of acetone were added to a 250mL three-necked flask and dissolved with stirring. Adding potassium carbonate (8g, 6.0mmol) and methyl iodide (12g, 0.8mol) under nitrogen protection, heating and refluxing for 24h, adding methyl iodide (4g, 0.3mol), refluxing for 12h, heating and concentrating to remove part of solvent, cooling to 25 ℃, adding 150mL of water, stirring for 8h to generate precipitate, filtering the precipitate, washing with water, and drying to obtain R-1(4.5g, 95%). 150mL of dried diethyl ether and tetramethylethylenediamine (3.1g, 27.0mmol) were charged in a dry 250mL three-necked flask, nitrogen gas was introduced, and a 1.6mol/L n-butyllithium-n-hexane solution (17mL,28.0mmol) was further added thereto, followed by stirring at room temperature for 30 minutes, R-1(3g, 9.5mmol) was further added thereto, and the mixture was stirred at room temperature for 3 hours. Adding triethyl borate slowly at-78 deg.C, heating to room temperature after adding, stirring for 24h, cooling to 0 deg.C, adding 75mL of 1mol/L HCl, stirring for 2h, washing the organic layer with 1mol/L HCl and saturated sodium chloride solution in sequence, and concentrating under reduced pressure to obtain R-2(3.1g, 65%).

Step two: a50 mL two-necked flask was charged with R-2(750mg, 1.9mmol), Ba (OH) 2.8H ₂ O (1.7g, 5.5mmol) and Pd (PPh) ₃ ) ₄ Introducing nitrogen, adding 12mL of 1, 4-dioxane, 4mL of water and 2-bromonaphthalene (4.5g, 6.0mmol), heating and refluxing for 24h under the protection of nitrogen, removing dioxane, and dissolving the crude product in 75mL of CH ₂ Cl ₂ The reaction mixture was washed with 1mol/L HCl (2X 50m L) and 75mL of saturated sodium chloride solution and dried over anhydrous sodium sulfate. Dissolve the crude in 75mL dry CH ₂ Cl ₂ In, cooled to 0 deg.C, 1mL BBr was added ₃ The mixture was stirred at room temperature for 18 hours. Cooled to 0 ℃ again, and 150mL of water was carefully added. The organic phase was separated, washed with water (2X 100mL) and 100mL of saturated sodium chloride solution, and dried over anhydrous sodium sulfateDrying, and concentrating under reduced pressure. The residue was purified by column chromatography (n-hexane: dichloromethane ═ 1:1, V/V) to give R-3(1.8g, 70%).

Step three: r-3(300mg, 0.43mmol) was added to a 10mL round-bottom flask, 1mL of pyridine was added and dissolved, and phosphorus oxychloride (0.081mL, 0.86mmol) was added dropwise at room temperature and stirred rapidly for 6 hours. 1mL of water was added and the resulting biphasic suspension was stirred at room temperature for 30 min. Dichloromethane was added and the organic phase was separated and washed with 1mol/L HCl (2X 20m L) and 25mL of saturated sodium chloride solution and dried over anhydrous sodium sulfate. Concentrating under reduced pressure. The residue was purified by column chromatography (methanol: dichloromethane ═ 1:90, V/V) to give BN (280mg, 86%). And (3) product identification result: ¹ H NMR(600MHz,CDCl ₃ )δ7.40-7.44(m,2H),7.46(d,J＝8.4Hz,1H),7.51(d,J＝9.6Hz,1H),7.53-7.55(m,2H),7.56-7.58(m,2H),7.60-7.63(m,2H),7.85(dd,J＝9.6Hz,1H),7.90-7.92(m,2H),7.93-7.96(m,3H),7.99(t,J＝7.8Hz,2H),8.09(t,J＝18.6Hz,2H),8.23(t,J＝18.6Hz,3H),8.28(s,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ122.81,123.06,126.22,126.36,126.55,126.73,127.10,127.19,127.64,127.75,127.96,128.21,128.45,128.69,129.17,131.97,132.08,132.22,132.41,132.94,133.40,133.52,133.79,134.00,144.27. ³¹ P NMR(243MHz,CDCl ₃ )δ3.60.MS(ESI-)m/z:599.1[M-H] ^- 。

example 17

The preparation method of diethyl aminomalonate described in this example:

to a 50mL round bottom flask was added diethyl aminomalonate hydrochloride (3.0g, 14.2mmol), followed by 30mL water dissolution and then NaHCO ₃ (1.3g, 15.6mmol), after stirring at room temperature for 15min, extraction was done 3 times with AcOEt, the organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum to give diethyl aminomalonate (2.3g, 13.1mmol) without further purification. And (3) product identification result: MS (ESI) M/z 198.0[ M + Na ]] ⁺ 。

Example 18

The preparation method of 1a described in this example:

in a 10mL round bottom flask were added (E) -1-acetyl-3-benzylideneindol-2-one (60mg, 0.2mmol), BN (10mg, 0.02mmol), compound A (47mg, 0.2mmol), 4A molecular sieves (300mg) and 2mL dry dichloromethane with stirring at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, and the reaction mixture was stirred at 25 ℃ for 24-48h, after completion of the reaction, and purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to give 1a (20mg, 20%). And (3) product result identification: m.p.69-71 ℃;

(c＝0.2in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.73(t,J＝7.1Hz,3H),1.34(t,J＝7.2Hz,3H),1.90-1.99(m,1H),2.59(s,3H),2.79(ddd,J＝2.7,8.4,16.6Hz,1H),3.03(t,J＝8.5Hz,1H),3.20(q,J＝8.4Hz,1H),3.64(s,3H),3.71(m,5H),4.35(q,J＝7.2Hz,1H),4.45(d,J＝8.3Hz,1H),4.51(d,J＝11.0Hz,1H),4.98(d,J＝11.0Hz,1H),5.60(s,1H),6.80(d,J＝7.4Hz,2H),7.00(t,J＝7.6Hz,2H),7.06(t,J＝7.4Hz,1H),7.19(td,J＝0.9,7.5Hz,1H),7.24(m,1H),7.46(d,7.3Hz,1H),7.53(s,1H),7.96(d,J＝8.1Hz,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.99,14.20,13.13,26.53,31.44,36.02,39.13,46.75,51.19,57.30,58.07,58.66,60.39,61.64,62.53,64.99,78.53,103.28,110.91,116.07,123.83,124.81,127.19,127.80,128.03,128.11,128.93,129.17,139.83,142.26,152.16,167.78,168.59,169.98,170.32,175.66.HRMS(ESI):Calcd.for C ₃₆ H ₃₉ O ₁₀ N ₂ [M+H] ⁺ :659.2599；found:659.2595。

example 19

The preparation method of 1b described in this example:

preparation method As in example 18, (E) -1-acetyl-6-chloro-3- (3 (trifluoromethyl) benzylidene) indol-2-one (66.2mg, 0.2mmol), BN (10mg, 0.02mmol), compound A (47mg, 0.2mmol), 4A molecular sieve (300mg), and 2mL of dried methylene chloride were added to a 10mL round bottom flask and stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, and the reaction mixture was stirred at 25 ℃ for 24-48h, after completion of the reaction, and purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to give 1b (22mg, 25%). And (3) product result identification: m.p.69-70 ℃;

(c＝0.3in CHCl ₃ )； ¹ HNMR(600MHz,CDCl ₃ )δ0.84(t,J＝7.1Hz,3H),1.32(t,J＝7.1Hz,3H),1.93(dd,J＝6.3,16.8Hz,1H),2.48(s,3H),2.68(dd,J＝7.9,16.6Hz,1H),2.98(t,J＝7.1Hz,1H),3.05(q,J＝7.8Hz,1H),3.54(s,3H),3.70(s,3H),4.13(dq,J＝7.0,14.2Hz,1H),4.25-4.34(m,2H),4.40(dq,J＝7.2,14.3Hz,1H),4.52(d,J＝7.0Hz,1H),4.70(d,J＝11.8Hz,1H),5.06(s,1H),5.19(s,1H),7.18(d,J＝7.6Hz,1H),7.26(t,J＝7.7Hz,1H),7.29(s,1H),7.41(s,1H),7.42-7.49(m,3H),8.10(s,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.23,13.96,26.34,34.68,39.08,47.11,51.19,56.68,61.99,62.84,63.66,64.27,66.83,74.86,101.72,110.57,117.18,122.59,124.88,125.60,125.90,125.97,128.77,130.70,133.89,134.71,134.96,137.25,140.47,151.96,167.52,176.86,170.16,170.45,176.31.HRMS(ESI):Calcd.for C ₃₇ H ₃₇ O ₁₀ N ₂ ClF ₃ [M+H] ⁺ :761.2083；found:761.2086。

example 20

The preparation method of 1c described in this example:

preparation method As in example 18, (E) -1-acetyl-6-chloro-3- (3- (methoxy) benzylidene) indol-2-one (65.4mg, 0.2mmol), BN (10mg, 0.02mmol), compound A (47.3mg,0.2mmol), 4A molecular sieves (300mg) and 2mL dry dichloromethane were stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, and the reaction mixture was stirred at 25 ℃ for 24-48h, after completion of the reaction, and purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to give 1c (25mg, 31%). And (3) product result identification: m.p.65-66 ℃;

(c＝0.28in CHCl ₃ )； ¹ HNMR(600MHz,CDCl ₃ )δ0.83(t,J＝7.1Hz,3H),1.33(t,J＝7.2Hz,3H),1.96(ddd,J＝1.6,9.2,16.7Hz,1H),2.60(s,3H),2.77-2.83(m,1H),3.01(t,J＝8.0Hz,1H),3.20(q,J＝8.4Hz,1H),3.60(s,3H),3.64(s,3H),3.72(s,3H),3.76-3.82(m,1H),3.83-3.92(m,1H),4.33(q,J＝7.2Hz,2H),4.43(s,1H),4.45(d,J＝4.5Hz,1H),4.90(d,J＝11Hz,1H),5.63(s,1H),6.33(s,1H),6.41(d,J＝7.7Hz,1H),6.64(dd,J＝2.0,8.2Hz,1H),6.97(t,J＝8.0Hz,1H),7.20(dd,J＝1.9,8.2Hz,1H),7.39(d,J＝8.2Hz,1H),7.53(s,1H),8.07(d,J＝1.8Hz,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.27,13.97,26.45,29.70,36.15,39.11,46.26,51.23,56.06,57.35,57.84,58.65,61.81,62.66,64.55,78.55,103.25,110.87,113.25,114.04,116.71,120.59,124.80,125.85,129.18,129.50,134.69,134.87,140.67,141.99,152.19,159.27,167.76,168.36,169.79,170.19,175.35.HRMS(ESI):Calcd.for C ₃₇ H ₄₀ O ₁₁ N ₂ Cl[M+H] ⁺ :723.2315；found:723.2311。

example 21

The preparation method of 1d described in this example:

preparation method As in example 18, (E) -1-acetyl-3- (3-chloro-2-fluorobenzylidene) -6- (trifluoromethyl) indol-2-one (81.4mg, 0.2mmol), BN (10mg, 0.02mmol), compound A (47.3mg, 0.2mmol), 4A molecular sieve (300mg), and 2mL dry methylene chloride are added to a 10mL round-bottomed flask and stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, and the mixture was reactedThe mixture was stirred at 25 ℃ for 24-48h and after completion of the reaction, it was purified by column chromatography (eluent: petroleum ether: ethyl acetate 5:1, V/V) to give 1d (26mg, 26%). And (3) product result identification: m.p.58-60 ℃;

(c＝0.12in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.86(t,J＝7.1Hz,3H),1.30(t,J＝7.2Hz,3H),1.99-2.06(m,1H),2.69(s,3H),2.76-2.87(m,1H),2.99(t,J＝7.8Hz,1H),3.21(q,J＝8.1Hz,1H),3.66(s,3H),3.73(s,3H),3.81-3.89(m,1H),3.90-4.00(m,1H),4.25-4.36(m,2H),4.55(d,J＝8.1Hz,1H),4.72(d,J＝10.7Hz,1H),5.15(d,J＝10.6Hz,1H),5.75(s,1H),6.73-6.91(m,2H),7.14-7.19(m,1H),7.22(d,J＝8.0Hz,1H),7.30(d,J＝8.3Hz,1H),7.54(s,1H),8.41(s,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.27,13.85,26.32,27.22,29.33,36.20,38.95,46.11,51.09,51.27,57.27,59.50,62.10,62.81,63.68,80.21,102.77,110.83,113.27,121.06,122.67,123.55,124.14,124.83,126.74,129.99,130.17,131.03,131.25,140.72,141.14,152.15,167.66.HRMS(ESI):Calcd.for C ₃₇ H ₃₆ O ₁₀ N ₂ ClF ₄ [M+H] ⁺ :779.1989；found:779.1982。

example 22

The preparation method of 1e described in this example:

preparation method As in example 18, in a 10mL round-bottom flask were added (E) -1-acetyl-6-chloro-3- (pyridin-4-ylmethylene) indol-2-one (60.0mg, 0.2mmol), BN (10mg, 0.02mmol), Ea (47.3mg, 0.2mmol), 4A molecular sieves (300mg), and 2mL dry dichloromethane, and stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, and the reaction mixture was stirred at 25 ℃ for 24-48h, after completion of the reaction, and purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to give 1e (27mg, 28%). And (3) product result identification: m.p.75-76 ℃;

(c＝0.34in CHCl ₃ )；1H NMR(600MHz,CDCl ₃ )δ0.84(t,J＝7.2Hz,3H),1.32(t,J＝7.1Hz,3H),1.74(s,2H),1.86–1.80(m,2H),1.98–1.87(m,2H),2.45–2.38(m,1H),2.60(s,3H),2.84–2.78(m,1H)),3.02(dd,J＝11.8,6.9Hz,1H),3.44(s,3H),3.51–3.47(m,2H),3.68–3.59(m,1H),3.70(d,J＝5.2Hz,1H),4.06–3.99(m,1H),4.31–4.25(m,1H),4.38(d,J＝4.1Hz,1H),4.47–4.39(m,1H),4.64(d,J＝4.7Hz,1H),5.09(s,1H),6.29(s,1H),6.82(d,J＝6.1Hz,2H),7.26(dd,J＝8.2,2.0Hz,1H),8.11(d,J＝1.9Hz,1H),8.33–8.30(m,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.37,14.00,26.68,27.21,29.32,33.90,34.17,35.92,37.23,37.45,49.06,56.37,60.44,62.09,62.91,63.79,64.96,73.26,99.14,115.62,116.82,123.95,124.57,124.79,128.58,132.54,134.73,137.26,141.10,142.31,142.84,149.48,167.74,168.87,170.42,170.50,176.94.HRMS(ESI):Calcd.for C ₃₈ H ₄₂ O ₉ N ₄ Cl[M+H] ⁺ :733.2635；found:733.2629。

example 23

The preparation method of 1f described in this example:

preparation method As in example 18, in a 10mL round-bottom flask were added (E) -1-acetyl-6-chloro-3- (pyridin-4-ylmethylene) indol-2-one (60.0mg, 0.2mmol), BN (10mg, 0.02mmol), Ga (47.3mg, 0.2mmol), 4A molecular sieve (300mg), and 2mL dry dichloromethane, and stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, and the reaction mixture was stirred at 25 ℃ for 24-48h, after completion of the reaction, and purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to give 1f (27mg, 28%). And (3) product result identification: m.p.102-103 ℃;

(c＝0.36in CHCl ₃ )； ¹ HNMR(600MHz,CDCl ₃ )δ0.83(t,J＝7.2Hz,3H),1.32(t,J＝7.1Hz,3H),2.41(dd,J＝7.1,16.4Hz,1H),2.60(s,3H),2.85(br.s.,1H),2.98-3.03(m,1H),3.17(br.s.,2H),3.43(s,3H),3.45-3.52(m,2H),3.53-3.58(m,2H),3.59-3.66(m,4H),3.97-4.07(m,1H),4.24-4.32(m,1H),4.37(d,J＝5.0Hz,1H),4.40-4.46(m,1H),4.70(d,J＝4.3Hz,1H),5.09(s,1H),5.72(s,1H),6.81(d,J＝6Hz,2H),7.25(dd,J＝1.9,8.2Hz,1H),8.10(d,J＝1.9Hz,1H),8.30-8.32(m,3H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.37,14.00,26.66,33.92,37.06,49.30,56.37,60.40,62.14,62.95,63.78,64.91,66.99,73.19,98.93,113.28,116.75,123.96,124.47,124.75,128.66,132.55,134.69,137.29,141.13,141.57,142.70,149.57,168.50,168.84,170.44,170.47,176.82.HRMS(ESI):Calcd.for C ₃₈ H ₄₂ O ₁₀ N ₄ Cl[M+H] ⁺ :749.2584；found:749.2579。

example 24

Preparation of 1g as described in this example:

preparation method As in example 18, in a 10mL round bottom flask were added (E) -1-acetyl-6-chloro-3- (3- (trifluoromethoxy) benzylidene) indol-2-one (60.0mg, 0.2mmol), BN (10mg, 0.02mmol), Compound A (47.3mg, 0.2mmol), 4A molecular sieves (300mg), and 2mL dry dichloromethane, and stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, and the reaction mixture was stirred at 25 ℃ for 24-48h, after completion of the reaction, and purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1g (27mg, 28%). And (3) product result identification: m.p.51-53 deg.C;

(c＝0.20in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.83(t,J＝7.1Hz,3H),1.33(t,J＝7.2Hz,3H),1.97(ddd,J＝1.7,9.2,16.8Hz,1H),2.60(s,3H),2.81(ddd,2.8,8.4,16.8Hz,1H),3.00(t,J＝7.3Hz,1H),3.21(q,J＝8.2Hz,1H),3.64(s,3H),3.72(s,3H),3.77-3.83(m,1H),3.83-3.90(m,1H),4.34(q,J＝7.1Hz,2H),4.43(d,J＝8.3Hz,1H),4.49(d,J＝11.0Hz,1H),4.93(d,J＝10.8Hz,1H),5.63(s,1H),6.69(s,1H),6.78(d,J＝7.9Hz,1H),6.98(d,J＝8.1Hz,1H),7.11(t,J＝8.0Hz,1H),7.20(dd,J＝1.9,8.2Hz,1H),7.37(d,J＝8.2Hz,1H),7.54(s,1H),8.07(d,J＝1.9Hz,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.27,13.96,26.41,29.70,36.23,39.06,46.24,51.25,57.25,57.28,58.56,61.90,62.78,64.44,78.57,103.13,110.83,116.81,120.64,120.76,124.67,125.04,125.13,126.61,129.67,129.87,135.10,135.93,140.49,141.64,148.94,152.20,167.70,168.16,169.67,170.04,175.21.HRMS(ESI):Calcd.for C ₃₇ H ₃₆ ClF ₃ N ₂ O ₁₁ [M+H] ⁺ :777.2032；found:777.2014。

example 25

The preparation method for 1h described in this example:

a10 mL round bottom flask was charged with tert-butyl (E) -4- ((1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) benzoate (79.0mg, 0.2mmol), BN (10mg, 0.02mmol), Compound A (47.3mg, 0.2mmol), 4A molecular sieves (300mg), and 2mL dry methylene chloride with stirring at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, the reaction mixture was stirred at 25 ℃ for 24-48h, and after completion of the reaction, purification was performed by column chromatography (eluent: petroleum ether: ethyl acetate ═ 5:1, V/V) to obtain 8, and compound 8 was dissolved in DCM, followed by addition of 1mL TFA, stirring at room temperature for 3h, concentration under vacuum, and purification on silica gel chromatography (petroleum ether: ethyl acetate: formic acid ═ 2:1:0.1) to obtain 1h (28mg, 33%). And (3) product result identification: m.p.92-94 ℃;

(c＝0.33in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.84(t,J＝7.1Hz,3H),1.33(t,J＝6.3Hz,3H),1.95(ddd,J＝1.7,9.2,16.8Hz,1H),2.75-2.81(m,1H),3.01(t,J＝7.4Hz,1H),3.19(q,J＝8.1Hz,1H),3.64(s,3H),3.72(s,3H),3.75-3.82(m,1H),3.83-3.90(m,1H),4.34(q,J＝7.1Hz,2H),4.43(d,J＝8.3Hz,1H),4.53(d,J＝11.0Hz,1H),5.00(d,J＝11.0Hz,1H),5.61(s,1H),6.93(d,J＝8.4Hz,2H),7.20(dd,J＝1.9,8.2Hz,1H),7.41(d,J＝8.2Hz,1H),7.53(s,1H),7.78(d,J＝8.3Hz,2H),8.06(d,1.9Hz,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.27,13.96,26.49,36.23,39.03,46.18,51.28,57.36,57.64,58.81,61.96,62.70,62.83,64.45,78.82,103.09,110.80,116.86,124.86,125.06,125.13,128.31,130.06,130.07,135.08,139.62,140.58,141.65,167.74,168.13,169.60,170.03,170.56,175.13.HRMS(ESI):Calcd.for C ₃₇ H ₃₈ ClN ₂ O ₁₂ [M+H] ⁺ :737.2；found:737.2086。

example 26

The preparation method of 1i described in this example:

into a 10mL round bottom flask was added (E) -1-acetyl-5-chloro-3- (3-chloro-2-fluorobenzylidene) indol-2-one (69.8mg, 0.2mmol), BN (10mg, 0.02mmol), Compound A (47.3mg, 0.2mmol), 4A molecular sieves (300mg), and 2mL dry dichloromethane with stirring at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, and the reaction mixture was stirred at 25 ℃ for 24-48h, after completion of the reaction, and purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to give 1i (26mg, 28%). And (3) product result identification: m.p.54-55 deg.C;

(c＝0.26in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.91(t,J＝7.1Hz,3H),1.29(d,J＝7.2Hz,3H),2.05–1.99(m,1H),2.66(s,3H),2.82(ddd,J＝16.8,8.4,2.8Hz,1H),2.99(t,J＝7.5Hz,1H),3.20(q,J＝8.2Hz,1H),3.68(s,3H),3.73(s,3H),4.01–3.90(m,2H),4.34–4.24(m,2H),4.54(d,J＝8.1Hz,1H),4.68(d,J＝10.7Hz,1H),5.16(d,J＝10.6Hz,1H),5.73(s,1H),6.93–6.85(m,2H),7.05(d,J＝2.1Hz,1H),7.17(td,J＝7.4,1.7Hz,1H),7.20(dd,J＝8.7,2.2Hz,1H),7.54(s,1H),8.05(d,J＝8.7Hz,1H). ¹³ C NMR(150MHz,CDCl ₃ )δ13.39,13.83,26.32,36.03,38.98,46.14,50.97,51.25,57.23,59.44,62.15,62.71,63.63,80.16,102.79,110.83,117.32,124.18,124.75,126.81,127.79,128.99,129.81,130.02,130.86,138.84,141.10,152.14,167.68,168.13,168.70,170.40,176.75.HRMS(ESI):Calcd.for C ₃₆ H ₃₆ Cl ₂ FN ₂ O ₁₀ [M+H] ⁺ :745.1726；found:745.1711。

example 27

The preparation method of 1j described in this example:

a10 mL round bottom flask was charged with (E) -1-acetyl-3- (3- (trifluoromethyl) benzylidene) indol-2-one (66.8mg, 0.2mmol), BN (10mg, 0.02mmol), Compound A (47.3mg, 0.2mmol), 4A molecular sieve (300mg), and 2mL dry dichloromethane and stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, and the reaction mixture was stirred at 25 ℃ for 24 to 48 hours, after completion of the reaction, and then purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to give 1j (23mg, 17%). And (3) product result identification: m.p.52-53 ℃;

(c＝0.56in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.82(t,J＝7.1Hz,3H),1.31(t,J＝7.1Hz,3H),1.96(ddd,J＝1.8,9.2,16.8Hz,1H),2.59(s,3H),2.75-2.82(m,1H),2.97-3.01(m,1H),3.19(q,J＝8.2Hz,1H),3.62(s,3H),3.71(s,3H),3.76-3.82(m,1H),3.76-3.82(m,1H),4.30-4.35(m,2H),4.42(d,J＝8.4Hz,1H),4.47(d,J＝11.0Hz,1H),4.91(d,J＝10.9Hz,1H),5.61(s,1H),6.68(s,1H),6.77(d,J＝7.7Hz,1H),6.96(d,J＝8.2Hz,1H),7.10(t,J＝8.0Hz,1H),7.19(dd,J＝1.9,8.2Hz,1H),7.36(d,J＝8.2Hz,1H),7.52(s,1H),8.06(d,J＝1.9Hz,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.23,13.96,26.34,34.68,39.08,47.11,51.19,56.68,61.99,62.84,63.66,64.27,66.83,74.86,101.72,110.57,117.18,122.59,124.88,125.60,125.90,125.97,128.77,130.70,133.89,134.71,134.96,137.25,140.47,151.96,167.52,176.86,170.16,170.45,176.31.HRMS(ESI):Calcd.for C ₃₇ H ₃₈ F ₃ N ₂ O ₁₀ [M+H] ⁺ :727.2473；found:727.2486。

example 28

The preparation method of 1k described in this example:

in a 10mL round bottom flask were added (E) -1-acetyl-6-chloro-3- (4- (trifluoromethyl) benzylidene) indol-2-one (73.1mg, 0.2mmol), BN (10mg, 0.02mmol), Compound A (47.3mg, 0.2mmol), 4A molecular sieves (300mg), and 2mL dry dichloromethane with stirring at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, and the reaction mixture was stirred at 25 ℃ for 48 hours, after completion of the reaction, and then purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1k (23mg, 25%). And (3) product result identification: m.p.66-68 ℃;

(c＝0.28in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.85(t,J＝7.1Hz,3H),1.33(t,J＝7.2Hz,3H),1.97(ddd,J＝16.8,9.2,1.8Hz,1H),2.61(s,3H),2.80(ddd,J＝16.8,8.2,2.9Hz,1H),3.02(dd,J＝11.4,4.6Hz,1H),3.20(q,J＝8.1Hz,1H),3.64(s,3H),3.73(s,3H),3.82–3.74(m,1H),3.93–3.83(m,1H),4.37–4.31(m,2H),4.43(d,J＝8.3Hz,1H),4.52(d,J＝11.0Hz,1H),4.96(d,J＝10.9Hz,1H),5.59(s,1H),6.96(d,J＝8.2Hz,2H),7.21(dd,J＝8.2,2.0Hz,1H),7.34(d,J＝8.2Hz,2H),7.41(d,J＝8.2Hz,1H),7.53(s,1H),8.10(d,J＝1.9Hz,1H). ¹³ C NMR(150MHz,CDCl ₃ )δ13.29,13.97,26.51,36.26,39.05,46.19,51.27,57.33,57.34,59.00,61.93,62.79,64.33,78.90,103.07,110.84,116.93,124.88,125.06,125.10,125.29,128.62,129.97,135.14,137.78,140.69,141.73,152.18,167.67,168.10,169.60,170.03,175.15.HRMS(ESI):Calcd.for C ₃₇ H ₃₇ ClF ₃ N ₂ O ₁₀ [M+H] ⁺ :761.2088；found:761.2061。

example 29

The preparation of 1l described in this example:

in a 10mL round bottom flask was added (E) -1-acetyl-6-chloro-3- (3-methoxy-5- (trifluoromethyl) benzylidene) indol-2-one (73.1mg, 0.2mmol), BN (10mg, 0.02mmol), Compound A (47.3mg, 0.2mmol), 4A molecular sieves (300mg), and 2mL dry dichloromethane with stirring at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, and the reaction mixture was stirred at 25 ℃ for 48 hours, after completion of the reaction, and then purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1l (20mg, 15%). And (3) product result identification: m.p.56-58 ℃;

(c＝0.37in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.84(t,J＝7.1Hz,1H),1.33(t,J＝7.2Hz,1H),2.02–1.95(m,1H),2.60(s,1H)),2.83(ddd,J＝16.7,8.4,2.8Hz,1H),2.99(t,J＝8.1Hz,1H),3.22(q,J＝8.2Hz,1H),3.63(s,1H),3.73(s,1H),3.90–3.80(m,1H),4.35(q,J＝7.2Hz,1H),4.43(d,J＝8.3Hz,1H),4.50(d,J＝11.0Hz,1H),4.91(d,J＝11.2Hz,1H),5.64(s,1H),6.50(s,1H),6.67(s,1H),6.87(s,1H),7.23(dd,J＝8.2,1.9Hz,1H),7.41(d,J＝8.2Hz,1H),7.54(s,1H). ¹³ C NMR(150MHz,CDCl ₃ )δ13.28,13.97,14.13,26.36,29.70,36.23,39.06,46.26,51.26,55.43,57.29,57.32,58.35,61.91,62.81,64.43,78.26,103.16,109.86,110.83,116.87,117.09,117.63,124.67,124.97,125.25,129.76,135.11,136.14,140.47,141.56,152.21,159.48,167.70,168.17,169.78,170.06,175.05.HRMS(ESI):Calcd.for C ₃₈ H ₃₉ ClF ₃ N ₂ O ₁₁ [M+H] ⁺ :791.2189；found:791.2172。

example 30

The preparation method of 1m described in this example:

dissolve 1a in 6mL EtOH/CH ₂ Cl ₂ To the mixed solution of 5/1, 60 μ L of 1M NaOH solution was added, stirred at room temperature for 2h, and then water was added. Extraction was performed twice with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1, V/V) to give 1m (20mg, 16%). And (3) product result identification: m.p.69-71 ℃;

(c＝0.34in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.77(t,J＝7.1Hz,3H),1.32(t,J＝7.2Hz,3H),1.93(ddd,J＝1.6,9.0,16.7Hz,1H),2.78(ddd,J＝2.7,8.4,16.8Hz,1H),3.03(t,J＝7.5Hz,1H),3.20(q,J＝8.3Hz,1H),3.64(s,3H),3.71(s,3H),3.75-3.85(m,2H),4.27-4.38(m,2H),4.46(d,J＝2.2Hz,1H),4.48(s,1H),4.99(d,J＝11.0Hz,1H),5.61(s,1H),6.60(d,J＝7.7Hz,1H),6.89(d,J＝7.5Hz,2H),7.01(td,2.8,7.5Hz,3H),7.06(t,J＝7.2Hz,1H),7.12(t,J＝7.7Hz,1H),7.38(d,J＝7.5Hz,1H),7.53(s,1H). ¹³ C NMR(150MHz,CDCl ₃ )δ13.18,13.94,35.93,39.09,46.38,51.20,53.44,56.91,57.28,58.54,61.49,62.38,64.91,77.81,103.29,109.39,110.95,122.17,124.56,127.30,127.76,128.52,128.64,129.08,134.10,140.53,142.37,152.18,167.86,168.97,170.21,175.96.HRMS(ESI):Calcd.for C ₃₄ H ₃₇ N ₂ O ₉ [M+H] ⁺ :617.2494；found:617.2479。

example 31

The preparation method of 1n described in this example:

to a 10mL round bottom flask was added methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) -1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] E]Pyran-4-carboxylic acid ester (85.8mg, 0.2mmol), BN (10mg, 0.02mmol), 2-fluoro-3-chlorobenzaldehyde (55.3mg, 0.2 mm)ol), 4A molecular sieves (300mg) and 2mL dry dichloromethane were stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, the reaction mixture was stirred at 50 ℃ for 48 hours, and after completion of the reaction, the mixture was purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1nn, and the 1nn was dissolved in 6mL of ethanol/CH ₂ Cl ₂ To the mixed solution of 5/1, 60 μ L of 1M NaOH solution was added, stirred at room temperature for 2h, and then water was added. Extracted twice with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1, V/V) to give 1n (25mg, 18%). And (3) product result identification: m.p.107-109 ℃;

(c＝0.21in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.89(t,J＝7.1Hz,3H),1.33(t,J＝7.2Hz,3H),1.68(t,J＝7.3Hz,1H),1.77-1.85(m,1H),2.53-2.65(m,2H),3.16(s,3H),3.67(s,3H),3.81-3.95(m,2H),4.14(d,J＝7.2Hz,1H),4.28-4.36(m,2H),4.46(d,J＝10.9Hz,1H),5.23(d,J＝10.9Hz,1H),5.28(s,1H),6.93(d,J＝1.8Hz,1H),7.07(dd,J＝1.8,8.1Hz,1H),7.18(t,J＝7.9Hz,1H),7.32(s,1H),7.34(d,J＝8.1Hz,1H),7.98(t,J＝6.5Hz,1H),8.28(s,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.32,13.89,34.95,39.18,47.98,51.17,52.32,56.66,58.14,60.43,61.94,62.58,63.18,77.51,102.17,110.30,110.46,120.90,122.47,124.86,125.71,127.04,127.53,129.69,130.41,133.17,134.92,136.45,142.48,151.76,167.66,168.39,170.04,175.04.HRMS(ESI):Calcd.for C ₃₄ H ₃₄ Cl ₂ FN ₂ O ₉ [M+H] ⁺ :703.1620；found:703.1602。

example 32

Preparation of 1o as described in this example:

to a 10mL round bottom flask was added methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] radical]Pyran-4-carboxylate (85.8mg, 0.2mmol), BN (10mg, 0.02mmol), 4-trifluoromethylbenzaldehyde (57.3mg, 0.2mmol), 4A molecular sieve (300mg) and 2mL dry methylene chloride were stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, the reaction mixture was stirred at 50 ℃ for 48 hours, after completion of the reaction, the reaction mixture was purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1oo, and the 1oo was dissolved in 6mL of ethanol/CH ₂ Cl ₂ To the mixed solution of 5/1, 60 μ L of 1M NaOH solution was added, stirred at room temperature for 2h, and then water was added. Extracted twice with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1, V/V) to give 1o (26mg, 18%). And (3) product result identification: m.p.114-116 ℃;

(c＝0.18in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.88(t,J＝7.1Hz,3H),1.36(t,J＝5.8Hz,3H),1.56(t,J＝7.1Hz,1H),1.77-1.82(m,1H),2.55-2.65(m,2H),3.26(s,3H),3.66(s,3H),3.84-3.94(m,2H),4.17(d,J＝7.3Hz,1H),4.32-4.38(m,3H),4.78(d,J＝10.8Hz,1H),5.06(s,1H),6.93(d,J＝1.8Hz,1H),7.06(dd,J＝1.8,8.1Hz,1H),7.30(s,1H),7.31(d,J＝8.2Hz,1H),7.62(d,J＝8.2Hz,2H),7.83(d,J＝8.1Hz,2H),8.53(br.s.,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.31,13.89,34.22,39.38,48.67,51.17,53.46,56.85,60.44,61.97,62.66,62.97,67.28,77.87,102.05,110.38,122.27,123.25,125.05,125.43,125.63,127.84,128.05,129.96,130.17,131.99,134.91,137.03,142.71,145.46,151.75,167.66,168.66,170.11,175.18.HRMS(ESI):Calcd.for C ₃₅ H ₃₅ ClF ₃ N ₂ O ₉ [M+H] ⁺ :719.1978；found:719.1960。

example 33

The preparation method of 1p described in this example:

to a 10mL round bottom flask was added methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) -1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] E]Pyran-4-carboxylate (85.8mg, 0.2mmol), BN (10mg, 0.02mmol), 3-trifluoromethylbenzaldehyde (57.3mg, 0.2mmol), 4A molecular sieve (300mg) and 2mL dry methylene chloride were stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, the reaction mixture was stirred at 50 ℃ for 48 hours, after completion of the reaction, the reaction mixture was purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1pp, and 1pp was dissolved in 6mL of a mixed solution of ethanol/CH 2Cl 2: 5/1, followed by addition of 60 μ L of a 1M NaOH solution, stirring at room temperature for 2 hours, and then water was added. Extraction was performed twice with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1, V/V) to give 1p (25mg, 18%). And (3) product result identification: m.p.97-99 ℃;

(c＝0.25in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.88(t,J＝7.1Hz,3H),1.36(t,J＝7.2Hz,3H),1.53(t,J＝7.3Hz,1H),1.81(dd,J＝6.4,15.4Hz,1H),2.55-2.65(m,2H),3.27(s,3H),3.66(s,3H),3.82-3.94(m,2H),4.19(d,J＝7.2Hz,1H),4.33(d,J＝11Hz,1H),4.34-4.41(m,2H),4.79(d,J＝10.8Hz,1H),5.06(s,1H),6.93(d,J＝1.8Hz,1H),7.06(dd,J＝1.8,8.1Hz,1H),7.30(t,4.0Hz,2H),7.49(t,J＝7.7Hz,1H),7.57(d,J＝7.8Hz,1H),7.89(d,J＝7.8Hz,1H),7.97(s,1H),8.39(s,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.30,13.80,34.12,39.38,48.70,51.15,53.30,56.80,60.44,62.01,62.66,62.91,67.23,77.85,101.97,110.38,122.26,123.28,124.69,125.09,125.56,127.85,128.90,130.64,130.89,132.12,134.93,136.92,142.54,142.66,151.72,167.64,168.69,170.00,175.04.HRMS(ESI):Calcd.for C ₃₅ H ₃₅ O ₉ N ₂ ClF ₃ [M+H] ⁺ :719.1978；found:719.1973。

example 34

The preparation method of 1q described in this example:

to a 10mL round bottom flask was added methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) -1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] E]Pyran-4-carboxylate (85.8mg, 0.2mmol), BN (10mg, 0.02mmol), 3-trifluoromethoxybenzaldehyde (57.3mg, 0.2mmol), 4A molecular sieve (300mg) and 2mL dry methylene chloride were stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, the reaction mixture was stirred at 50 ℃ for 48 hours, after completion of the reaction, the reaction mixture was purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1qq, 1qq was dissolved in 6mL of a mixed solution of ethanol/CH 2Cl 2: 5/1, 60 μ L of 1M NaOH solution was added, and the mixture was stirred at room temperature for 2 hours, followed by addition of water. Extraction was performed twice with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1, V/V) to give 1q (24mg, 18%). And (3) product result identification: m.p.82-84 ℃;

(c＝0.32in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.89(t,J＝7.1Hz,3H),1.37(t,J＝7.2Hz,3H),1.56(t,J＝7.4Hz,1H),1.80(dd,J＝7.3,16.3Hz,1H),2.54-2.61(m,1H),2.63(dd,J＝7.8,15.6Hz,1H),3.28(s,3H),3.66(s,3H),3.81-3.96(m,2H),4.21(d,J＝7.1Hz,1H),4.30(d,J＝10.8Hz,1H),4.32-4.41(m,2H),4.73(d,J＝10.8Hz,1H),5.02(s,1H),6.93(d,J＝1.8Hz,1H),7.07(dd,J＝1.8,8.1Hz,1H),7.16(d,J＝8.2Hz,1H),7.29(s,1H),7.31(s,1H),7.39(t,J＝8.1Hz,1H),7.60-7.64(m,2H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.32,13.82,34.01,39.36,48.70,51.13,53.63,56.81,61.96,62.67,62.83,67.20,77.83,101.95,110.29,110.44,119.66,120.36,121.36,122.29,125.61,125.97,127.90,129.79,131.92,134.89,136.91,142.57,143.94,149.43,151.72,167.66,168.70,170.03,173.78.HRMS(ESI):Calcd.for C ₃₅ H ₃₅ ClF ₃ N ₂ O ₁₀ [M+H] ⁺ :735.1927；found:735.1910。

example 35

The preparation method of 1r described in this example:

to a 10mL round bottom flask was added methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) -1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] E]Pyran-4-carboxylate (85.8mg, 0.2mmol), BN (10mg, 0.02mmol), 3-fluoro-4-chlorobenzaldehyde (49.3mg, 0.2mmol), 4A molecular sieve (300mg) and 2mL dry dichloromethane were stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added, the reaction mixture was stirred at 50 ℃ for 48 hours, after completion of the reaction, the mixture was purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1rr, and the 1rr was dissolved in 6mL of ethanol/CH ₂ Cl ₂ To the mixed solution of 5/1, 60 μ L of 1M NaOH solution was added, stirred at room temperature for 2h, and then water was added. Extraction was performed twice with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1, V/V) to give 1r (25mg, 18%). And (3) product result identification: m.p.102-104 ℃;

(c＝0.36in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.88(t,J＝7.1Hz,3H),1.35(t,J＝7.2Hz,3H),1.59(t,J＝7.4Hz,1H),1.81(dd,J＝7.4,16.4Hz,1H),2.60(ddd,J＝2.5,8.0,11.1Hz,1H),2.65(dd,J＝7.8,15.6Hz,1H),3.31(s,3H),3.67(s,3H),3.82-3.93(m,2H),4.22(d,J＝7.2Hz,1H),4.30(d,J＝10.7Hz,1H),4.32-4.39(m,2H),4.77(d,J＝10.7Hz,1H),5.07(s,1H),6.93(d,J＝1.8Hz,1H),7.07(dd,J＝1.8,8.1Hz,1H),7.27(d,J＝8.1,1H),7.32(s,1H),7.50(d,J＝8.3Hz,1H),7.82(dd,J＝1.7,8.3Hz,1H),8.07(d,J＝1.6Hz,1H),8.22(brs.,1H). ¹³ CNMR(151MHz,CDCl ₃ )δ13.29,13.81,34.18,39.36,48.80,51.18,53.69,56.81,60.43,62.06,62.69,62.83,66.57,77.73,101.85,110.41,121.99,122.29,123.80,125.48,127.07,127.64,128.23,131.48,131.95,132.56,135.03,136.68,140.90,142.61,151.71,167.59,168.56,169.87,174.80.HRMS(ESI):Calcd.for C ₃₅ H ₃₄ Cl ₂ F ₃ N ₂ O ₉ [M+H] ⁺ :753.1568；found:753.1562。

example 36

The preparation method of 1s described in this example:

to a 10mL round bottom flask was added methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) -1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] E]Pyran-4-carboxylate (85.8mg, 0.2mmol), BN (10mg, 0.02mmol), 4-chloro-3-fluoro-benzaldehyde (69.3mg, 0.2mmol), 4A molecular sieve (300mg) and 2mL dry dichloromethane were stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, the reaction mixture was stirred at 50 ℃ for 48 hours, after completion of the reaction, the mixture was purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1ss, and the 1ss was dissolved in 6mL of ethanol/CH ₂ Cl ₂ To the mixed solution of 5/1, 60 μ L of 1M NaOH solution was added, stirred at room temperature for 2h, and then water was added. Extraction was performed twice with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1, V/V) to give 1s (21mg, 14%). And (3) product result identification: m.p.102-104 ℃;

(c＝0.36in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.88(t,J＝7.1Hz,3H),1.35(t,J＝7.2Hz,3H),1.59(t,J＝7.4Hz,1H),1.81(dd,J＝7.4,16.4Hz,1H),2.60(ddd,J＝2.5,8.0,11.1Hz,1H),2.65(dd,J＝7.8,15.6Hz,1H),3.31(s,3H),3.67(s,3H),3.82-3.93(m,2H),4.22(d,J＝7.2Hz,1H),4.30(d,J＝10.7Hz,1H),4.32-4.39(m,2H),4.77(d,J＝10.7Hz,1H),5.07(s,1H),6.93(d,J＝1.8Hz,1H),7.07(dd,J＝1.8,8.1Hz,1H),7.27(d,J＝8.1,1H),7.32(s,1H),7.50(d,J＝8.3Hz,1H),7.82(dd,J＝1.7,8.3Hz,1H),8.07(d,J＝1.6Hz,1H),8.22(brs.,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.29,13.81,34.18,39.36,48.80,51.18,53.69,56.81,60.43,62.06,62.69,62.83,66.57,77.73,101.85,110.41,121.99,122.29,123.80,125.48,127.07,127.64,128.23,131.48,131.95,132.56,135.03,136.68,140.90,142.61,151.71,167.59,168.56,169.87,174.80.HRMS(ESI):Calcd.for C ₃₅ H ₃₄ C _l2 F ₃ N ₂ O ₉ [M+H] ⁺ :753.1568；found:753.1562。

example 37

The preparation method of 1t described in this example:

to a 10mL round bottom flask was added methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) -1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] E]Pyran-4-carboxylate (85.8mg, 0.2mmol), BN (10mg, 0.02mmol), 2-trifluoromethylbenzaldehyde (48.3mg, 0.2mmol), 4A molecular sieves (300mg) and 2mL dry methylene chloride were stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, the reaction mixture was stirred at 50 ℃ for 48 hours, after completion of the reaction, the mixture was purified by column chromatography (eluent: petroleum ether: ethyl acetate ═ 5:1, V/V) to obtain 1tt, the 1tt was dissolved in 6mL of a mixed solution of ethanol/CH 2Cl2 ═ 5/1, 60 μ L of a 1M NaOH solution was added, and the mixture was stirred at room temperature for 2 hours, followed by addition of water. Extraction was performed twice with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1, V/V) to give 1t (22mg, 15%). And (3) product result identification: m.p.105-106 ℃;

(c＝0.53in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.9(t,J＝7.1Hz,3H),1.41(t,J＝7.2Hz,3H),2.44-2.52(m,1H),2.54(dd,J＝8.0,16.0Hz,1H),3.45(s,3H),3.63(s,3H),3.78-3.86(m,1H),3.87-3.95(m,1H),4.17(d,J＝7.5Hz,1H),4.37(q,J＝7.1Hz,2H),4.60(d,J＝10.8Hz,1H),4.76(s,1H),5.09(d,J＝10.9Hz,1H),6.95(d,J＝1.8Hz,1H),7.11(dd,J＝1.9,8.1Hz,1H),7.24(d,J＝8.1Hz,1H),7.41(t,J＝7.6Hz,1H),7.62(d,J＝7.7Hz,1H),7.70(t,J＝7.6Hz,1H),8.68(d,J＝8.0Hz,1H),8.82(s,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.31,13.91,33.50,39.49,48.33,51.14,52.85,57.27,60.47,61.74,62.03,62.51,62.67,77.76,102.64,110.48,122.64,123.49,125.30,125.43,128.04,128.92.129.12,130.52,130.68,132.63,134.91,135.76,139.66,142.75,151.64,167.76,168.53,169.79,175.68.HRMS(ESI):Calcd.for C ₃₅ H ₃₅ ClF ₃ N ₂ O ₉ [M+H] ⁺ :719.1978；found:719.1962。

example 38

The preparation method of 1u described in this example:

to a 10mL round bottom flask was added methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) -1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] E]Pyran-4-carboxylate (85.8mg, 0.2mmol), BN (10mg, 0.02mmol), 3-bromobenzaldehyde (48.3mg, 0.2mmol), 4A molecular sieve (300mg) and 2mL dry methylene chloride were stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, the reaction mixture was stirred at 50 ℃ for 48 hours, and after completion of the reaction, the mixture was purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1uu, which was dissolved in 6mL of ethanol/CH ₂ Cl ₂ To the mixed solution of 5/1, 60 μ L of 1M NaOH solution was added, stirred at room temperature for 2h, and then water was added. Extraction was performed twice with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1, V/V) to give 1u (23mg, 15%). And (3) product result identification: m.p.87-89 ℃;

(c＝0.21in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.88(t,J＝7.1Hz,3H),1.41(t,J＝7.2Hz,3H),1.59(t,J＝7.5Hz,1H),1.78-1.83(m,1H),2.59(ddd,J＝2.6,8.2,16.7,1H),2.64(dd,J＝7.8,15.6Hz,1H),3.30(s,3H),3.67(s,3H),3.83-3.94(m,2H),4.20(d,J＝7.3Hz,1H),4.29(d,J＝10.8Hz,1H),4.38(q,J＝7.2Hz,2H),4.68(d,J＝10.8Hz,1H),4.99(s,1H),6.93(d,J＝1.8Hz,1H),7.06(dd,J＝1.9,8.1Hz,1H),7.22(t,J＝7.8Hz,1H),7.28(s,1H),7.32(s,1H),7.43(dd,J＝0.9,7.9Hz,1H),7.54(d,J＝7.8Hz,1H),7.94(s,1H),8.35(s,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.32,13.94,34.12,39.42,48.75,51.18,53.57,56.94,61.99,62.72,62.87,67.34,77.85,102.08,110.36,110.44,122.26,122.67,125.58,126.13,127.92,129.98,130.78,131.02,131.80,134.87,137.03,142.60,143.70,151.77,167.71,168.72,170.04,175.11.HRMS(ESI):Calcd.for C ₃₄ H ₃₅ BrClN ₂ O ₉ [M+H] ⁺ :729.1209；found:729.1190。

example 39

Preparation of 1v described in this example:

to a 10mL round bottom flask was added methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) -1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] E]Pyran-4-carboxylate (85.8mg, 0.2mmol), BN (10mg, 0.02mmol), 3-chlorobenzaldehyde (48.3mg, 0.2mmol), 4A molecular sieve (300mg) and 2mL dry methylene chloride were stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, the reaction mixture was stirred at 50 ℃ for 48 hours, after completion of the reaction, the reaction mixture was purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1vv, the 1vv was dissolved in 6mL of a mixed solution of ethanol/CH 2Cl 2: 5/1, 60 μ L of 1M NaOH solution was added, and the mixture was stirred at room temperature for 2 hours, followed by addition of water. Extraction was performed twice with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate ═ 2:1, V/V) to give 1V (24mg, 14%). And (3) product result identification: m.p.104-105 ℃;

(c＝0.32in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.88(t,J＝7.1Hz,3H),1.41(t,J＝7.2Hz,3H),1.58(t,J＝7.5Hz,1H),1.79(dd,J＝7.4,16.5Hz,1H),2.59(ddd,J＝2.5,8.0,11.2Hz,1H),2.64(dd,J＝7.8,15.7Hz,1H),3.30(s,3H),3.66(s,3H),3.83-3.94(m,2H),4.19(d,J＝7.3Hz,1H),4.30(d,J＝10.8,1H),4.38(q,J＝7.2Hz,2H),4.68(d,J＝10.8Hz,1H),4.99(s,1H),6.93(d,J＝1.8Hz,1H),7.06(dd,J＝1.8,8.1Hz,1H),7.27(s,1H),7.29(s,1H),7.31(s,1H),7.47-7.51(m,1H),7.80(s,1H),8.37(s,1H). ¹³ C NMR(151MHz,CDCl ₃ )δ13.32,13.91,34.12,39.43,48.73,51.18,53.46,53.57,56.94,60.46,61.98,62.72,62.87,67.43,77.88,102.11,110.35,110.43,122.26,125.59,125.71,127.86,127.94,128.10,129.67,131.71,134.45,134.87,137.07,142.63,143.45,151.76,167.70,168.73,170.07,175.14.HRMS(ESI):Calcd.for C ₃₄ H ₃₅ Cl ₂ N ₂ O ₉ [M+H] ⁺ :685.1714；found:685.1699。

example 40

The preparation method of 1w described in this example:

to a 10mL round bottom flask was added methyl (1R) -7- (((E) -1-acetyl-6-chloro-2-oxoindol-3-ylidene) methyl) -1-methoxy-1, 4a,5,7 a-tetrahydrocyclopenta [ c ] E]Pyran-4-carboxylate (85.8mg, 0.2mmol), BN (10mg, 0.02mmol), 3-fluorobenzaldehyde (45.3mg, 0.2mmol), 4A molecular sieve (300mg), and 2mL dry methylene chloride were stirred at 25 ℃. Diethyl aminomalonate (35mg, 0.2mmol) was added thereto, the reaction mixture was stirred at 50 ℃ for 48 hours, and after completion of the reaction, the mixture was purified by column chromatography (eluent: petroleum ether: ethyl acetate: 5:1, V/V) to obtain 1ww, and 1ww was dissolved in 6mL of a mixed solution of ethanol/CH 2Cl 2: 5/1, and 60 μ L of a 1M NaOH solution was added thereto, stirred at room temperature for 2 hours, and then water was added thereto. Extracting with dichloromethane twice, washing with saturated sodium chloride solution, drying with anhydrous sodium sulfate, and reducing pressureConcentration gave a crude product which was purified by silica gel column chromatography (petroleum ether: ethyl acetate 2:1, V/V) to give 1w (21mg, 15%). And (3) product result identification: m.p.96-98 deg.c;

(c＝0.31in CHCl ₃ )； ¹ H NMR(600MHz,CDCl ₃ )δ0.88(t,J＝7.1Hz,3H),1.39(t,J＝7.2Hz,3H),1.56(t,J＝7.5Hz,1H),1.78(dd,J＝7.4,16.1Hz,1H),2.58(ddd,J＝2.5,8.0,11.2Hz,1H),2.62(dd,J＝7.9,15.7Hz,1H),3.29(s,3H),3.66(s,3H),3.83-3.93(m,2H),4.18(d,J＝7.3Hz,1H),4.30(d,J＝10.8Hz,1H),4.37(q,J＝7.2Hz,2H),4.69(d,J＝10.8Hz,1H),4.96(s,1H),6.92(d,J＝1.8Hz,1H),6.99(td,J＝2.1,8.2Hz,1H),7.06(dd,J＝1.8,8.1Hz,1H),7.30(s,2H),7.35(d,J＝7.7Hz,1H),7.58(d,J＝10.0Hz,1H),8.53(d,J＝2.7Hz,1H). ¹³ CNMR(151MHz,CDCl ₃ )δ13.31,13.88,34.08,39.44,48.67,51.17,53.59,56.93,60.46,61.96,62.70,62.89,67.54,77.89,102.17,110.35,110.40,114.50,114.66,114.83,114.97,122.25,123.27,125.60,127.99,129.81,129.85,131.47,134.84,137.16,142.71,144.02,144.07,151.76,162.65,163.88,167.71,168.75,170.14,171.28,175.25.HRMS(ESI):Calcd.for C ₃₄ H ₃₅ ClFN ₂ O ₉ [M+H] ⁺ :669.2010；found:669.1994。

EXAMPLE 41 test of inhibitory Activity of the Compound of the present invention against 3 tumor cells

The MTT method is adopted, and the inhibition effect of the compound on the growth of three tumor cell strains in vitro is respectively tested by taking human non-small cell lung cancer HCC827, human highly metastatic liver cancer cell MHCC97H and human neuroblastoma SHSY-5Y as screening objects. The specific experimental steps are as follows:

1. inoculating cells: a single cell suspension was prepared using a culture medium (DMEM or RMPI1640) containing 10% fetal bovine serum, and 96 wells were seeded with 3000 to 15000 cells per well.

2. Adding a solution of the test compound: compounds were dissolved in DMSO and 3 replicates were provided for each treatment.

3. Color development: after culturing at 37 ℃ for 48 hours, the cells were discarded from the culture medium in the wells, and MTT solution and culture medium were added to each well. And continuously incubating for 2-4 hours to ensure that the light absorption value is measured after the reaction is fully carried out.

4. Color comparison: and selecting the wavelength of 492nm, reading the light absorption value of each hole by a multifunctional microplate reader, and recording the result. IC50 was calculated using an IC50 calculator. Results the results shown in table 1 are shown in table 1:

TABLE 1 test results of antitumor Activity of the Compounds of the present invention in vitro

As can be seen from Table 1, most of the compounds have certain inhibitory action on human non-small cell lung cancer HCC827, human highly metastatic liver cancer cell MHCC97H and human neuroblastoma SHSY-5Y, and have inhibitory activity on three tumor cells better than genipin, especially have strong inhibitory action on human neuroblastoma SHSY-5Y,

in particular 1g, 1j, 1l of compound. The compounds are proved to have stronger selectivity on neuroblastoma SHSY-5Y. The compound containing trifluoromethyl has stronger inhibition effect on three tumor cells, such as compound 1g, 1i, 1j, 1k and 1 l. Shows that the trifluoromethyl has important function on the antitumor activity of the compound.

Finally, it should be noted that: although the present invention has been described in detail with reference to the above embodiments, it should be understood by those skilled in the art that: modifications and equivalents may be made thereto without departing from the spirit and scope of the invention and it is intended to cover in the claims the invention as defined in the appended claims.

Claims

1. A spirocyclic indolone compound based on iridoid aglycone shown in formula I,

wherein R is selected from H, Cl; r ₁ Selected from H, Cl, F, trifluoromethyl, trifluoromethoxy; r ₂ Is methyl; r is ₃ Selected from acetyl; x is CH.

2. A spirocyclic indolone compound based on iridoid aglycon shown in a formula II,

wherein R is selected from trifluoromethoxy.

3. The iridoid aglycone-based spiro indolone compound is characterized by comprising the following components in percentage by weight:

4. a process for the preparation of a compound according to claim 1, wherein a compound of formula (iii-1) is reacted with a compound of formula (a) or (b) to produce a compound of formula (i):

wherein R is selected from H, Cl; r ₁ Selected from H, Cl, F, trifluoromethyl, trifluoromethoxy; r ₂ Is methyl; r ₃ Selected from acetyl; x is CH; the BN structural formula is

5. A process for the preparation of a compound according to claim 2, wherein a compound of formula (iii-2) is reacted with a compound of formula (c) or (b) to form a compound of formula (ii):

wherein R is selected from trifluoromethoxy; the BN structural formula is

6. The use of iridoid aglycon-based spirocyclic indolones according to any one of claims 1 to 3 for the preparation of a medicament against resistant tumors.