A benzazepine spiroindoline compound and its synthesis method and application

Abstract

The present invention relates to the technical field of organic compound synthesis, and particularly relates to a benzazepine spirodihydroindole compound, a synthesis method and an application thereof. A tryptamine derivative, an o-iodobenzoic acid or o-bromobenzoic acid derivative, and an isonitrile derivative are successively added to a methanol solution of an aldehyde or ketone compound, and the mixture is stirred at room temperature overnight. The product precipitates in solid form, and the Ugi adduct is obtained by filtration. The Ugi adduct, Pd(OAc)2, Ph3P, and K2CO3 are added to a sealed container and stirred. The container is evacuated and filled with nitrogen, and anhydrous o-dichlorobenzene is added and the reaction is sealed. The reaction is monitored by TLC, and the sample is loaded wet and subjected to silica gel column chromatography to obtain the target product. Starting from cheap and readily available common raw materials, the present invention rapidly combines aldehydes, amines, carboxylic acids, and isonitriles into Ugi adducts with diverse skeletons and high functionality through the Ugi four-component reaction under mild reaction conditions. Using this as a substrate, a highly chemoselective dearomatization cyclization reaction of a complex Ugi adduct catalyzed by palladium is carried out to obtain a series of benzazepine spirodihydroindole derivatives.

Description

Technical Field

[0001] The invention relates to the technical field of organic compound synthesis, and in particular to a benzazepine spiroindoline compound and a synthesis method and application thereof. Background Art

[0002] Natural product structural frameworks provide biologically pre-validated and evolutionarily selected starting points for drug development in a vast chemical structure space. A new synthetic method suitable for the rapid construction of compound collections is expected to address the synthetic challenges posed by natural products and analogs. Tryptamine-derived natural products and their analogs, such as vinblastine, physostigmine, etc., are a large class of indole alkaloids with remarkable biological properties and daunting structural diversity. They have become the benchmark for the development of selective methods to coordinate concise bond-forming chemistry, thus stimulating the interest of countless chemists as total synthetic targets. Although a large amount of synthetic work has been done on these natural products, there is still a high demand for synthetic methods that can provide a large number of natural products and their analogs with different skeletons for biological activity testing.

[0003] Currently, the products obtained in reports of indole dearomatization are generally limited to 5- or 6-membered spiroindoline derivatives. Catalytic dearomatization of indoles and their derivatives has played a key role in the rapid assembly of structurally complex nitrogen-containing polycyclic frameworks by destroying the aromatic π system. Notably, transition metal-catalyzed dearomatization, especially Pd-catalyzed cross-coupling dearomatization, represents an attractive atom-economical approach for highly chemo- and stereo-selective dearomatization to generate polycyclic systems. However, the application of this strategy to structurally complex molecules containing multiple nucleophilic sites, especially in addressing the main problem, i.e., the collective and chemo-selective synthesis of bioactive molecules, remains underexplored. This may be due to the high reactivity of the in situ generated Pd(II) oxidative addition complexes, while the multiple nucleophilic sites in the substrates are difficult to control, resulting in poor chemoselectivity. In addition, the cumbersome preparation of Pd-catalyzed dearomatization substrates also hinders its application. To overcome the great challenges in achieving high levels of chemoselectivity, examples of sensitive functional groups involved in previous dearomatization cases have been limited to at most two free NH or OH groups, such as aniline, phenol, tryptamine, tryptophan, N-(2-halobenzoyl)indole, and indoles with pendant anilines or carbon nucleophiles. In addition, when there is no substituent at the 2- or 3-position of the electron-rich indole, it is easily captured by electron-deficient Pd(II) complexes to generate Friedel-Crafts-type compounds, which is a common chemoselectivity problem in the current Pd-catalyzed dearomatization of indoles. In this context, it is of great theoretical significance and practical prospect to study and develop a cost-effective and sustainable method to precisely control the dearomatization pathway to reproduce the architectural diversity and complexity of natural products. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide a benzazepine spiroindoline compound and a synthesis method and application thereof, starting from cheap and readily available common raw materials, through the Ugi four-component reaction, aldehydes, amines, carboxylic acids, and isonitriles are quickly kneaded into skeleton-diverse, highly functionalized Ugi adducts under mild reaction conditions, and using these as substrates through a palladium-catalyzed highly chemically selective dearomatization cyclization reaction of the complex Ugi adducts to obtain a series of benzazepine spiroindoline derivatives inspired by natural products.

[0005] The present invention solves the above technical problems by the following technical means:

[0006] The first aspect of the present invention is to provide a benzazepine spiroindoline compound, characterized in that the structural formula of the benzazepine spiroindoline compound is as follows:

[0007]

[0008] Among them, R1 is alkyl, halogen or hydrogen, R2 is alkyl, aryl or heteroaryl, R3 is alkyl or aryl, and Ar is aryl or heteroaryl.

[0009] The second aspect of the present invention is to provide a method for synthesizing a benzazepine spiroindoline compound, comprising the following steps:

[0010] S1. Compound 2, compound 3 and compound 4 are sequentially added to a methanol solution containing compound 1, mixed and stirred, precipitated, and filtered to obtain a Ugi adduct, wherein compound 1 is an aldehyde or ketone compound, compound 2 is a tryptamine derivative, compound 3 is an o-iodobenzoic acid or o-bromobenzoic acid derivative, and compound 4 is an isonitrile derivative;

[0011] S2. Add Ugi adduct, Pd(OAc)2, Ph3P and K2CO3 into a sealed container and stir to mix, evacuate the container and fill it with nitrogen, repeat this operation three times, then add anhydrous 1,2-dichlorobenzoate and seal the reaction, monitor by TLC, and perform wet loading and silica gel column chromatography on the obtained reaction mixture to obtain the target product benzazepine spirodihydroindole compound;

[0012] The synthetic route is as follows:

[0013]

[0014] Among them, R1 is alkyl, halogen or hydrogen, R2 is alkyl, aryl or heteroaryl, R3 is alkyl or aryl, and Ar is aryl or heteroaryl.

[0015] The third aspect of the present invention is to provide the use of the above-mentioned benzazepine spiroindoline compound or the benzazepine spiroindoline compound synthesized by the above-mentioned synthesis method in the preparation of anti-tumor drugs.

[0016] The synthesis method of the present invention can quickly obtain a series of benzazepine spiroindoline compounds. The synthesis method starts from cheap and easily available common raw materials, and quickly combines aldehydes, amines, carboxylic acids and isonitriles under mild reaction conditions to form Ugi adducts with diverse skeletons and high functionalization. The Ugi adducts are used as substrates to carry out a highly chemically selective dearomatization cyclization reaction of complex Ugi adducts catalyzed by palladium, thereby obtaining a series of benzazepine spiroindoline derivatives inspired by natural products.

[0017] The synthesis method of the invention has excellent chemical selectivity and good diastereoselectivity. The combination of Ugi-4CR and palladium-catalyzed dearomatization is efficient and easy to operate, so a natural product compound library can be quickly synthesized for medicinal chemistry and chemical biology research. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 is the general structural formula of the benzazepine spiroindoline compound of the present invention;

[0019] Figure 2 is a synthetic route diagram of the benzazepine spiroindoline compound of the present invention;

[0020] Figure 3 This is a graph showing the anti-tumor activity test results of the benzazepine spiroindoline compounds synthesized in Examples 1-10;

[0021] Figure 4 This is a graph showing the effect of different concentrations of the benzazepine spiroindoline compound synthesized in Example 6 on the activity of breast cancer cells;

[0022] Figure 5 The IC values ​​of breast cancer cells treated with the benzazepine spiroindoline compounds synthesized in Example 6 at different concentrations are shown in Table 1. 50 Influence diagram;

[0023] Figure 6 is a statistical diagram of apoptosis of breast cancer cells treated with the benzazepine spiroindoline compounds synthesized in Example 6 at different concentrations;

[0024] Figure 7 is a flow cytometric plot;

[0025] Figure 8 It is the single crystal structure of the benzazepine spiroindoline compound synthesized in Example 1 when the TLC polarity position is the lower point;

[0026] Fig. 9It is the single crystal structure of the benzazepine spiroindoline compound synthesized in Example 1 when the TLC polarity position is the upper point;

[0027] Fig.10 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 1 when the TLC polarity position is the lower point;

[0028] Fig.11 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 1 when the TLC polarity position is the upper point;

[0029] Fig.12 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 2 when the TLC polarity position is the lower point;

[0030] Fig.13 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 2 when the TLC polarity position is the upper point

[0031] Fig.14 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 3 when the TLC polarity position is the lower point

[0032] Fig.15 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 3 when the TLC polarity position is the upper point

[0033] Fig.16 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 4 when the TLC polarity position is the lower point

[0034] Fig.17 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 5 when the TLC polarity position is the lower point

[0035] Fig.18 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 6 when the TLC polarity position is the lower point

[0036] Fig.19 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 7 when the TLC polarity position is the lower point

[0037] Fig. 20 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 8 when the TLC polarity position is the upper point

[0038] Fig.21 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 9 when the TLC polarity position is the lower point

[0039] Fig. 22This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 10 when the TLC polarity position is the lower point

[0040] Fig.23 This is the NMR spectrum of the benzazepine spiroindoline compound synthesized in Example 11 when the TLC polarity position is the lower point

[0041] Fig.24 This is the nuclear magnetic resonance spectrum of the benzazepine spiroindoline compound synthesized in Example 11 when the TLC polarity position is the upper point. DETAILED DESCRIPTION

[0042] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0043] The benzazepine spiroindoline compounds of the present invention have the following characteristics: Figure 1 The general structural formula shown, wherein R1 is alkyl, halogen or hydrogen, R2 is alkyl, aryl or heteroaryl, R3 is alkyl or aryl, Ar is aryl or heteroaryl, preferably R1 is hydrogen, 6-fluoro, 5-bromo, 5-methoxy, R2 is phenyl, 3,4,5-trimethoxyphenyl, 3-pyridyl, isopropyl, R3 is tert-butyl, 2,4,4-trimethylpentyl, Ar is phenyl, 4-trifluoromethylphenyl, 4,5-dimethoxyphenyl, thienyl.

[0044] The above-mentioned benzazepine spiroindoline compound is used as Figure 2 The synthetic route shown is synthesized, and the specific steps are as follows:

[0045] S1: To a solution of the aldehyde compound (3 mmol, 1.0 equiv.) in methanol (10 mL) were added a tryptamine derivative (3 mmol, 1.0 equiv.), an o-iodobenzoic acid derivative (3 mmol, 1.0 equiv.) and an isonitrile derivative (2.8 mmol, 0.93 equiv.) in sequence. The reaction mixture was stirred at room temperature overnight in a screw-capped vial equipped with a magnetic stirring bar. After the reaction was completed, the product precipitated in the form of a solid, which was filtered to directly obtain the desired Ugi adduct 1.

[0046] S2: Add Ugi adduct 1 (0.1mmol), Pd(OAc)2 (0.005mmol, 1.1mg, 0.05eq), Ph3P (0.01mmol, 2.6mg, 0.1eq) and K2CO3 (0.2mmol, 28mg, 2.0eq) to a dry Schlenk flask equipped with a stirring bar, evacuate the flask and fill it with nitrogen, and repeat this operation three times. Then, add anhydrous 1,2-dichlorobenzene (1mL) and seal the flask. The reaction mixture was stirred at 120°C for 16 hours and monitored by TLC. The reaction mixture was then wet-loaded and column chromatographed, and the target product, benzazepine spirodihydroindole compound, was obtained by silica gel column chromatography with ethyl acetate / n-hexane = 30% to 40% as the eluent.

[0047] The following Examples 1-11 all adopt the above preparation method to obtain the corresponding benzazepine spiroindoline compounds.

[0048] Example 1

[0049] In the benzazepine spiroindoline compound synthesized in this embodiment, R1 is hydrogen, R2 is phenyl, R3 is tert-butyl, and Ar is phenyl, that is, the benzazepine spiroindoline compound of this embodiment is N-(tert-butyl)-2-(1-oxo-3,4-dihydrospiro[benzo[c]azacyclo-5,3'-indole]-2(1H)-yl)-2-phenylacetamide, and its structural formula is as follows:

[0050]

[0051] The benzazepine spiroindoline compound of this example is a white solid when the TLC polarity position is at the lower point, and its structural formula is as follows Figure 8 As shown, the yield is 78%, and the ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf = 0.25 (ethyl acetate / hexane = 30%). Fig.10 shown.

[0052] 1 H NMR (400MHz, CDCl3) δ8.25(s,1H),7.92(d,J=7.6Hz,1H),7.65(d,J=7.4Hz,1H),7.52(d,J=7.5Hz,2H),7.46(d,J=7.2Hz,1H),7.42–7.35(m,5H) ,7.33–7.29(m,2H),7.05(d,J=7.8Hz,1H),6.34(s,1H),5.63(s,1H),3. 72–3.69(m,2H),2.10–2.05(m,1H),1.40(s,9H),1.09–1.01(m,1H)ppm;

[0053] 13 C NMR (100MHz, CDCl3) δ174.1,171.5,168.7,154.5,140.1,136.7,135.1,131.5,131.2,131.1,129. 8,129.3,129.1,128.6,128.4,126.4,125.6,125.0,122.0,64.5,61.2,52.0,43.2,40.8,28.7ppm;

[0054] HRMS(ESI)m / z calcd for C 29 H 30 N3O2 + (M+H) + 452.2333, found m / z 452.2337.

[0055] The benzazepine spiroindoline compound of this example is a yellow solid when the TLC polarity position is up, and its structural formula is as follows Fig. 9 As shown, the yield is 11%, and the ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf = 0.5 (ethyl acetate / hexane = 30%). Fig.11 shown.

[0056] 1 H NMR (400MHz, CDCl3) δ8.29(s,1H),7.89(d,J=7.2Hz,1H),7.70(d,J=6.3Hz,1H),7.52(d,J=5.6Hz,1H),7.40–7.31(m,9H),7.12(d,J =7.5Hz,1H),6.49(s,1H),6.05(s,1H),3.55–3.52(m,1H),3.39–3.33(m,1H),2.65–2.58(m,1H),2.27–2.23(m,1H),1.41(s,9H)ppm;

[0057] 13 C NMR (100MHz, CDCl3) δ174.7,171.9,167.9,154.8,140.3,136.7,135.6,132.4,131.5,130.4,1 29.0,128.5(t,J=8.8Hz),126.3,125.7,125.0,122.1,64.6,60.7,52.1,42.9,40.1,28.6ppm;

[0058] HRMS(ESI)m / z calcd for C29 H 30 N3O2 + (M+H) + 452.2333, found m / z 452.2335.

[0059] Example 2

[0060] In the benzazepine spiroindoline compound synthesized in this embodiment, R1 is hydrogen, R2 is 3,4,5-trimethoxyphenyl, R3 is tert-butyl, and Ar is phenyl. That is, the benzazepine spiroindoline compound of this embodiment is N-(tert-butyl)-2-(1-oxo-3,4-dihydrospiro[benzo[c]azacyclo-5,3'-indole]-2(1H)-yl)-2-(3,4,5-trimethoxyphenyl)acetamide, and its structural formula is as follows:

[0061]

[0062] The benzazepine spiroindoline compound of this example is a white solid when the TLC polarity position is the lower point, and the yield is 71%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf = 0.25 (ethyl acetate / hexane = 45%). Its NMR spectrum is as follows Fig.12 shown.

[0063] 1 H NMR (400MHz, CDCl3) δ8.36(s,1H),7.92(d,J=7.5Hz,1H),7.65(d,J=7.1Hz,1H),7.48(d,J=6.7Hz,1H),7.45–7.35(m,3H),7.30(t,J=7.6Hz, 1H),7.09(d,J=7.7Hz,1H),6.73(s,2H),6.24(s,1H),5.71(s,1H),3.8 2(s,9H),3.74–3.62(m,2H),2.13–2.06(m,1H),1.23–1.19(m,1H)ppm;

[0064] 13 C NMR (100MHz, CDCl3) δ174.1,171.6,168.8,154.5,153.5,140.0,138.2,136.6,131.6,131.2,131.1,130 .3,128.7,128.4,126.6,125.6,125.1,122.1,106.7,64.6,61.1,60.9,56.2,52.0,43.2,41.3,28.7ppm;

[0065] HRMS(ESI)m / z calcd for C 32 H 36 N3O5 + (M+H) + 542.2649, found m / z 542.2655.

[0066] The benzazepine spiroindoline compound of this example is a white solid when the TLC polarity position is the upper point, and the yield is 13%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf = 0.5 (ethyl acetate / hexane = 45%). Its NMR spectrum is as follows Fig.13 shown.

[0067] 1 H NMR (400MHz, CDCl3) δ8.28(s,1H),7.89(d,J=7.6Hz,1H),7.69(d,J=7.6Hz,1H),7 .54(d,J=6.8Hz,1H),7.45–7.36(m,3H),7.32(t,J=7.6Hz,1H),7.13(d,J=7.8Hz, 1H),6.66(s,2H),6.36(s,1H),6.01(s,1H),3.84(s,3H),3.78(s,6H),3.59–3.51 (m,1H),3.42–3.31(m,1H),2.69–2.58(m,1H),2.25–2.18(m,1H),1.42(s,9H)ppm;

[0068] 13 C NMR (100MHz, CDCl3) δ174.9,171.9,167.9,154.8,153.5,140.5,138.1,136.6,132.7,131.5,130.8,130 .4,128.6,128.3,126.3,125.7,124.9,122.1,105.9,64.5,61.1,60.9,56.2,52.0,42.8,39.9,28.6ppm;

[0069] HRMS(ESI)m / z calcd for C 32 H 36 N3O5 + (M+H) + 542.2649, found m / z 542.2651.

[0070] Example 3

[0071] In the benzazepine spiroindoline compound synthesized in this embodiment, R1 is hydrogen, R2 is 3-pyridyl, R3 is tert-butyl, and Ar is phenyl, that is, the benzazepine spiroindoline compound of this embodiment is N-(tert-butyl)-2-(1-oxo-3,4-dihydrospiro[benzo[c]azacyclo-5,3'-indole]-2(1H)-yl)-2-pyridin-3-yl)acetamide, and its structural formula is as follows:

[0072]

[0073] The benzazepine spiroindoline compound of this example is a yellow solid when the TLC polarity position is the lower point, and the yield is 63%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf = 0.25 (ethyl acetate / hexane = 40%). Its NMR spectrum is as follows Fig.14 shown.

[0074] 1 H NMR (400MHz, CDCl3) δ8.77(s,1H),8.58(d,J=4.2Hz,1H),8.22(s,1H),7.91(t,J=8.4Hz,2H),7.65(d,J=7.0Hz,1H),7.47(d,J=7.0Hz,1H),7 .44–7.30(m,5H),7.08(d,J=7.8Hz,1H),6.34(s,1H),6.01(s,1H),3.7 8–3.64(m,2H),2.16–2.13(m,1H),1.39(s,9H),1.20–1.12(m,1H)ppm;

[0075] 13 C NMR (100MHz, CDCl3) δ173.6,171.8,167.7,154.4,150.6,150.2,139.9,137.5,136.2,131.8,131. 4,131.1,128.7,128.5,126.6,125.7,125.0,123.9,122.2,64.4,59.2,52.1,43.2,40.7,28.7ppm;

[0076] HRMS(ESI)m / z calcd for C 28 H 29 N4O2 + (M+H) + 453.2285, found m / z 453.2286.

[0077] The benzazepine spiroindoline compound of this example is a yellow oil when the TLC polarity position is the upper point, and the yield is 21%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf=0.4 (ethyl acetate / hexane=40%). Its NMR spectrum is as follows Fig.15 shown.

[0078] 1 H NMR (400MHz, CDCl3) δ8.56(s,1H),8.50(s,1H),8.24(s,1H),7.83(d,J=7.6Hz,1H),7.68 (d,J=7.9Hz,1H),7.63(d,J=6.7Hz,1H),7.47(d,J=7.5Hz,1H),7.41–7.32(m,3H),7.29– 7.20(m,2H),7.06(d,J=7.8Hz,1H),6.60(s,1H),6.55(s,1H),3.56(dd,J=14.7,5.1Hz,1 H),3.40–3.29(m,1H),2.51–2.40(m,1H),2.25(dd,J=14.4,3.4Hz,1H),1.32(s,9H)ppm;

[0079] 13 C NMR (100MHz, CDCl3) δ173.2,171.0,166.0,153.6,148.7,148.4,139.0,135.2,135.0,131.1,130.7 ,129.4,127.7,127.4,125.50(s),124.8,124.0,122.6,121.1,63.5,57.1,51.3,41.8,39.0,27.5;

[0080] HRMS(ESI)m / z calcd for C 28 H 29 N4O2 + (M+H) + 453.2285, found m / z 453.2284.

[0081] Example 4

[0082] In the benzazepine spiroindoline compound synthesized in this embodiment, R1 is hydrogen, R2 is propyl, R3 is tert-butyl, and Ar is phenyl, that is, the benzazepine spiroindoline compound of this embodiment is N-(tert-butyl)-3-methyl-2-(1-oxo-3,4-dihydrospiro[benzo[c]azacyclo-5,3'-indole]-2(1H)-yl)butanamide, and its structural formula is as follows:

[0083]

[0084] The benzazepine spiroindoline compound of this example is a yellow solid when the TLC polarity position is the lower point, and the yield is 82%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf=0.4 (ethyl acetate / hexane=30%). Its NMR spectrum is as follows Fig.16 shown.

[0085] 1 H NMR(400MHz, CDCl3)δ8.20(s,1H),7.79(d,J=7.5Hz,1H),7.74–7.65(m,1H),7.61–7.50(m, 1H),7.47–7.37(m,3H),7.30(dd,J=10.8,4.5Hz,1H),7.10(d,J=7.8Hz,1H),6.35(s,1H),4 .82(d,J=10.9Hz,1H),3.88(dd,J=14.5,4.4Hz,1H),3.47(td,J=14.4,4.2Hz,1H),2.40–2. 30(m,2H),2.28–2.20(m,1H),1.34(s,9H),1.01(d,J=6.4Hz,3H),0.90(d,J=6.7Hz,3H)ppm;

[0086] 13 C NMR (100MHz, CDCl3) δ174.3,171.8,169.0,154.6,140.0,137.1,131.4,131.3,130.0,128. 7,128.3,126.6,125.6,125.1,122.1,64.3,62.7,51.9,41.1,39.9,28.5,27.5,19.2,18.3;

[0087] HRMS(ESI)m / z calcd for C 26 H 32 N3O2 + (M+H) + 418.2489, found m / z 418.2488.

[0088] Example 5

[0089] In the benzazepine spiroindoline compound synthesized in this embodiment, R1 is hydrogen, R2 is phenyl, R3 is tert-butyl, and Ar is methylphenyl, that is, the benzazepine spiroindoline compound of this embodiment is N-(tert-butyl)-2-(8-methyl-1-oxo-3,4-dihydrospiro[benzo[c]azacyclo-5,3'-indole]-2(1H)-yl)-2-phenylacetamide, and its structural formula is as follows:

[0090]

[0091] The benzazepine spiroindoline compound of this example is a white solid when the TLC polarity position is the lower point, and the yield is 83%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf=0.5 (ethyl acetate / hexane=40%). Its NMR spectrum is as follows Fig.17 shown.

[0092] 1 H NMR(400MHz, CDCl3)δ8.21(s,1H),7.65(d,J=7.4Hz,1H),7.51–7.46(m,4H),7.42–7.33(m,5H),6.51(s,1H),6.31(s, 1H),5.59(s,1H),3.94(s,3H),3.78–3.67(m,2H),3.64(s,3H),2.08–2.03(m,1H),1.40(s,9H),1.06–0.98(m,1H)ppm;

[0093] 13 C NMR (100MHz, CDCl3) δ174.5,171.4,168.8,154.6,150.9,148.5,140.6,135.3,129.9,129.2,129.1,12 9.0,128.6,126.4,124.5,124.4,122.2,114.0,109.0,64.3,61.4,56.1,55.7,52.0,43.4,40.9,28.8;

[0094] HRMS(ESI)m / z calcd for C 31 H 34 N3O4 + 512.2544, found m / z 512.2548.

[0095] Example 6

[0096] In the benzazepine spiroindoline compound synthesized in this embodiment, R1 is hydrogen, R2 is phenyl, R3 is tert-butyl, and Ar is trifluoromethylphenyl. That is, the benzazepine spiroindoline compound of this embodiment is N-(tert-butyl)-2-(1-oxo-7-(trifluoromethyl)-3,4-dihydrospiro[benzo[c]azacyclo-5,3'-indole]-2(1H)-yl)-2-phenylacetamide, and its structural formula is as follows:

[0097]

[0098] The benzazepine spiroindoline compound of this example is a yellow oil when the TLC polarity position is at the lower point, and the yield is 82%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf = 0.3 (ethyl acetate / hexane = 30%). Its NMR spectrum is as follows Fig.18 shown.

[0099] 1 H NMR (400MHz, CDCl3) δ8.24(s,1H),8.05(d,J=8.0Hz,1H),7.67(t,J=6.0Hz,2H),7.51(d,J=7.4Hz,2H),7.47–7.36(m,5H),7.33(d,J= 8.2Hz,2H),6.35(s,1H),5.59(s,1H),3.78–3.59(m,2H),2.08(dd,J=14.3,2.9Hz,1H),1.40(s,9H),1.06(td,J=13.7,5.9Hz,1H)ppm;

[0100] 13 C NMR (100MHz, CDCl3) δ173.0,170.2,168.4,154.4,140.0,139.2,134.7,133.1(q,J=32.7Hz),132.5, 131.7,129.8,129.4,129.3,129.0,127.0,125.2,124.7,122.4,64.3,61.2,52.1,43.0,40.8,28.7;

[0101] 19 F NMR (376MHz, CDCl3) δ-63.01 (t, J=5.7Hz).

[0102] HRMS(ESI)m / z calcd for C 30 H 29 F3N3O2 + (M+H) +520.2206, found m / z 520.2211.

[0103] Example 7

[0104] In the benzazepine spiroindoline compound synthesized in this embodiment, R1 is hydrogen, R2 is phenyl, R3 is tert-butyl, and Ar is thienyl, that is, the benzazepine spiroindoline compound of this embodiment is N-(tert-butyl)-2-(8'-oxo-5', 6'-dihydrospiro[indole-3,4'-thieno[2,3-c]azaheterocycle]-7'(8'H)-yl)-2-phenylacetamide, and its structural formula is as follows:

[0105]

[0106] The benzazepine spiroindoline compound of this example is a yellow solid when the TLC polarity position is the lower point, and the yield is 86%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf=0.3 (ethyl acetate / hexane=30%). Its NMR spectrum is as follows Fig.19 shown.

[0107] 1 H NMR(400MHz, CDCl3)δ7.96(s,1H),7.63(d,J=7.5Hz,1H),7.45–7.35(m,6H),7.31–7.22(m,3H),6.42(s,1H),6.08(d,J=5.2Hz,1H),5.75(s, 1H),3.94(dd,J=15.1,9.4Hz,1H),3.60(dd,J=15.2,6.8Hz,1H),2.20(dd,J=15.1,6.9Hz,1H),1.81(dd,J=14.7,9.3Hz,1H),1.40(s,9H)ppm;

[0108] 13 C NMR (100MHz, CDCl3) δ174.9,168.6,164.3,154.5,143.7,138.0,136.9,135.6,130.9, 129.2,129.1,128.7,128.3,127.2,123.0,121.6,63.3,61.8,52.0,43.1,35.0,28.7;

[0109] HRMS(ESI)m / z calcd for C 23 H 28 NO + (M+H) + 458.1897, found m / z 458.1899.

[0110] Example 8

[0111] In the benzazepine spiroindoline compound synthesized in this embodiment, R1 is a methoxy group, R2 is a phenyl group, R3 is a tert-butyl group, and Ar is a phenyl group, that is, the benzazepine spiroindoline compound of this embodiment is N-(tert-butyl)-2-(5'-methoxy-1-oxo-3,4-dihydrospiro[benzo[c]azacyclo-5,3'-indole]-2(1H)-yl)-2-phenylacetamide, and its structural formula is as follows:

[0112]

[0113] The benzazepine spiroindoline compound of this example is a yellow solid when the TLC polarity position is the lower point, and the yield is 78%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf=0.3 (ethyl acetate / hexane=30%). Its NMR spectrum is as follows Fig. 20 shown.

[0114] 1 H NMR (400MHz, CDCl3) δ8.17 (s, 1H), 7.89 (d, J = 7.5Hz, 1H), 7.61–7.58 (m, 1H), 7.41–7.31(m,7H),7.15(d,J=7.6Hz,1H),7.06(s,1H),6.95(d,J=8.4Hz,1H), 6.51(s,1H),6.15(s,1H),3.90(s,3H),3.58–3.53(m,1H),3.34(dd,J=19.2, 7.9Hz,1H),2.61(td,J=12.8,4.2Hz,1H),2.21–2.18(m,1H),1.40(s,9H)ppm;

[0115] 13 C NMR (100MHz, CDCl3) δ172.8,171.9,167.9,158.6,148.4,142.0,136.6,135.6,132.4,131.5,130.4 ,128.9,128.4,128.3,128.2,125.7,122.4,112.9,111.8,64.7,60.6,55.9,52.1,42.8,40.4,28.6;

[0116] HRMS(ESI)m / z calcd for C 30 H 32 N3O3 + (M+H) +482.2438, found m / z 482.2455.

[0117] Example 9

[0118] In the benzazepine spiroindoline compound synthesized in this embodiment, R1 is fluorine, R2 is phenyl, R3 is tert-butyl, and Ar is phenyl, that is, the benzazepine spiroindoline compound of this embodiment is N-(tert-butyl)-2-(6'-fluoro-1-oxo-3,4-dihydrospiro[benzo[c]azacyclo-5,3'-indole]-2(1H)-yl)-2-phenylacetamide, and its structural formula is as follows:

[0119]

[0120] The benzazepine spiroindoline compound of this example is a yellow solid when the TLC polarity position is the lower point, and the yield is 84%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf = 0.25 (ethyl acetate / hexane = 30%). Its NMR spectrum is as follows Fig.21 shown.

[0121] 1 H NMR (400MHz, CDCl3) δ8.31(s,1H),7.93(dd,J=7.7,1.3Hz,1H),7.51(d,J=7.1Hz,2H),7.43–7.37(m,3H),7.37–7.28(m,4H),7.10–7 .05(m,1H),7.02–7.00(m,1H),6.34(s,1H),5.62(s,1H),3.71–3.67(m,2H),2.06–2.01(m,1H),1.39(s,9H),1.08–1.00(m,1H)ppm;

[0122] 13 C NMR (100MHz, CDCl3) δ176.1, 171.4, 168.6, 164.3, 161.9, 155.98 (d, J = 10.9Hz), 136.6, 135.60 (d, J = 2.9Hz), 135.1, 131.6 ,131.2,130.9,129.8,129.3,129.1,128.5,125.6,125.4,113.4,113.2,109.8,109.5,64.2,61.1,52.0,43.1,40.7,28.7;

[0123] 19 F NMR(376MHz, CDCl3)δ-113.21–-113.27(m)

[0124] HRMS(ESI)m / z calcd for C 29 H 29 FN3O2 + (M+H) + 470.2238, found m / z 470.2244.

[0125] Example 10

[0126] In the benzazepine spiroindoline compound synthesized in this embodiment, R1 is bromine, R2 is phenyl, R3 is tert-butyl, and Ar is phenyl, that is, the benzazepine spiroindoline compound of this embodiment is N-(tert-butyl)-2-(6'bromo-1-oxo-3,4-dihydrospiro[benzo[c]azacyclo-5,3'-indole]-2(1H)-yl)-2-phenylacetamide, and its structural formula is as follows:

[0127]

[0128] The benzazepine spiroindoline compound of this example is a yellow solid when the TLC polarity position is the lower point, and the yield is 81%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf=0.25 (ethyl acetate / hexane=30%). Its NMR spectrum is as follows Fig. 22 shown.

[0129] 1 H NMR (400MHz, CDCl3) δ8.22(s,1H),7.93(dd,J=7.7,1.4Hz,1H),7.60(d,J=1.1Hz,1H),7.52–7.49(m,4H),7.46–7.43(m,1H),7.41–7.31( m,4H),7.02(dd,J=7.8,0.8Hz,1H),6.33(s,1H),5.62(s,1H),3.75–3.63(m,2H),2.06–2.01(m,1H),1.39(s,9H),1.09–1.00(m,1H)ppm;

[0130] 13 C NMR (100MHz, CDCl3) δ174.5,171.3,168.6,153.5,142.4,136.6,135.1,131.8,131.7,131.3,130 .5,129.8,129.3,129.1,128.6,128.3,125.4,123.4,120.3,65.0,61.1,52.0,43.1,40.7,28.7;

[0131] HRMS(ESI)m / z calcd for C29 H 29 BrN3O2 + (M+H) + 530.1438, found m / z 530.1443.

[0132] Embodiment 11

[0133] In the benzazepine spiroindoline compound synthesized in this embodiment, R1 is hydrogen, R2 is phenyl, R3 is 2,4,4-trimethylpentyl, and Ar is phenyl. That is, the benzazepine spiroindoline compound of this embodiment is 2-(1-oxo-3,4-dihydrospiro[benzo[c]azacyclo-5,3'-indol]-2(1H)-yl)-2-phenyl-N-(2,4,4-trimethylpentane-2-yl)acetamide, and its structural formula is as follows:

[0134]

[0135] The benzazepine spiroindoline compound of this example is a white solid when the TLC polarity position is the lower point, and the yield is 79%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf = 0.3 (ethyl acetate / hexane = 30%). Its NMR spectrum is as follows Fig.23 shown.

[0136] 1 H NMR(400MHz, CDCl3)δ8.15(s,1H),7.82(dd,J=7.7,1.2Hz,1H),7.59–7.54(m,1H),7.48(d,J=7 .2Hz,2H),7.41–7.36(m,1H),7.34–7.26(m,5H),7.22(dd,J=10.7,4.2Hz,2H),6.97(d,J=7.6Hz ,1H),6.26(s,1H),5.63(s,1H),3.76–3.41(m,2H),1.99(dd,J=14.4,3.0Hz,1H),1.85(d,J=14. 9Hz,1H),1.48(d,J=14.9Hz,1H),1.41(s,3H),1.36(s,3H),1.05–0.94(m,1H),0.91(s,9H)ppm;

[0137] 13C NMR (100MHz, CDCl3) δ174.1,171.1,168.2,154.6,140.1,136.8,134.9,131.4,131.3,131.0,130.0,129.2,1 29.0,128.5,128.3,126.4,125.6,125.0,122.0,64.5,61.2,55.9,52.4,43.11,40.7,31.6,31.5,29.2,28.5;

[0138] HRMS(ESI)m / z calcd for C 33 H 38 N3O2 + (M+H) + 508.2959, found m / z 508.2966.

[0139] The benzazepine spiroindoline compound of this example is a white solid when the TLC polarity position is the upper point, and the yield is 20%. The ratio of the distance of the substance moving on the silica gel chromatography plate to the distance of the solvent line is Rf=0.5 (ethyl acetate / hexane=30%). Its NMR spectrum is as follows Fig.24 shown.

[0140] 1 H NMR (400MHz, CDCl3) δ8.24(s,1H),7.80(dd,J=7.6,1.4Hz,1H),7.65–7.61(m,1H),7.44(dd,J=7.1,1. 1Hz,1H),7.35(ddd,J=5.5,4.9,1.8Hz,4H),7.32–7.27(m,4H),7.25–7.19(m,1H),7.04(dd,J=7.8,0. 8Hz,1H),6.38(s,1H),5.86(s,1H),3.45(ddd,J=14.7,5.4,1.8Hz,1H),3.36–3.21(m,1H),2.56(ddd, J=14.1,12.3,5.5Hz,1H),2.23–2.09(m,1H),1.64(s,2H),1.40(s,3H),1.38(s,3H),0.90(s,9H)ppm;

[0141] 13C NMR (100MHz, CDCl3) δ174.8,171.8,167.6,154.9,140.5,136.7,135.5,132.48,131.4,130.4,129.0,128.7,12 8.5,128.5,128.2,126.2,125.7,124.9,122.1,64.6,61.5,56.1,52.5,43.0,40.0,31.6,31.5,28.8,28.4ppm;

[0142] HRMS(ESI)m / z calcd for C 33 H 38 N3O2 + (M+H) + 508.2959, found m / z 508.2963.

[0143] Example 12 Antitumor Activity Test Experiment

[0144] The anti-tumor activity of the benzazepine spiroindoline compounds synthesized in Examples 1-10 was tested using human tumor cell lines A549, MCF7, HELA, HEPG2, and HCT116, all of which were purchased from the American Type Culture Collection (ATCC). The experimental steps are as follows:

[0145] (1) Routine cell culture:

[0146] A549, MCF7, HELA, HEPG2, and HCT116 cells were inoculated in DMEM medium containing 10% fetal bovine serum (FBS) and cultured and passaged at 37°C, 5% CO2, and saturated humidity. The cells were used for experiments when they were in the logarithmic growth phase;

[0147] (2) Cell plating

[0148] 24 h before drug treatment, A549, MCF7, HELA, HEPG2, and HCT116 cells in the logarithmic growth phase were digested and prepared into cell suspensions, which were counted and adjusted to a cell concentration of 1x10 5 / ml, the cells were seeded in a 96-well plate, with approximately 2x10 cells per well. 4 The cells were cultured in an incubator;

[0149] (3) Drug treatment

[0150] The benzazepine spiroindoline compounds synthesized in Example 1-10 were diluted with the complete culture medium of the corresponding cells to a compound concentration of 20 μM, and added to each well of a 96-well plate, with 3 replicates for each compound. The control group was treated with DMSO of the same volume as the compound, and the cells were returned to the cell culture incubator and cultured for 72 hours;

[0151] (4) CCK8 test results

[0152] 1) After the culture plate is placed in an incubator for 72 hours, the absorbance is detected at 450 nm using an ELISA reader;

[0153] 2) Add 10ul of CCK8 solution to each well (be careful not to create bubbles in the wells, as it will affect the reading);

[0154] 3) Place the culture plate in an incubator and incubate for 2 hours;

[0155] 4) Read the reading at 450nm, the results are as follows:

[0156] Table 1 shows the cell viability of the benzazepine spiroindoline compounds synthesized in Examples 1-10 on various tumor cells.

[0157] Table 1 Antitumor activity of benzazepine spiroindoline compounds

[0158]

[0159] Experimental results: The effects of 20 μM of the benzazepine spiroindoline compounds synthesized in Examples 1-10 on the growth ability of human non-small cell lung cancer cell line (A549), breast cancer cell line (MCF7), cervical cancer cell line (HELA), liver cancer cell line (HEPG2), and colon cancer cell line (HCT116) are as follows: Figure 3 shown. Figure 3 The data show that the benzazazepine spiroindoline compounds synthesized in Examples 1-10 have a certain inhibitory effect on the growth ability of breast cancer cells and colon cancer cells, especially the benzazazepine spiroindoline compound synthesized in Example 6 shows a good inhibitory effect on the growth ability of the tested breast cancer cells. The experimental results show that the benzazazepine spiroindoline synthesized in Example 6 can inhibit the growth of tumor cells, has good anti-tumor activity, and can be used to prepare anti-tumor drugs, especially the benzazazepine spiroindoline compound synthesized in Example 6 can be used to prepare anti-tumor drugs for treating breast cancer.

[0160] Example 13 Antitumor Activity and IC of the Benzazapine Spiroindoline Compounds Synthesized in Example 6 50 Test experiment

[0161] (1) Routine cell culture

[0162] In order to detect the effect of different concentrations of the benzazepine spiroindoline compound synthesized in Example 6 on the viability of breast cancer cells (MCF7), MCF7 cells were inoculated in DMEM culture medium containing 10% FBS, cultured and passaged at 37°C, 5% CO2 and saturated humidity, and the cells were used for experiments when they were in the logarithmic growth phase.

[0163] (2) Cell plating

[0164] 24 h before drug treatment, the MCF7 cells in the growth phase were digested and prepared into a cell suspension, which was counted and adjusted to a cell concentration of 1x10 5 / mL, the cells were seeded in a 96-well plate, with approximately 2x10 cells per well. 4 The cells were cultured in an incubator.

[0165] (3) Drug treatment

[0166] After the cells adhered, the benzazepine spirodihydroindole compound synthesized in Example 6 was added to give a final concentration of 0 μM, 1.25 μM, 2.5 μM, 5 μM, 10 μM, and 20 μM. After 72 hours of compound treatment, the absorbance was detected using an ELISA instrument at 450 nm. 10 ul of CCK solution was added to each well (be careful not to generate bubbles in the well), the culture plate was placed in an incubator and incubated for 2 hours, and then the reading was taken at 450 nm. The experimental results are shown in FIG. Figure 4 As shown in the figure, as the concentration of the compound increases, the survival ability of breast cancer cells is significantly reduced, indicating that the benzazepine spiroindoline compound synthesized in Example 6 has a certain inhibitory activity on breast cancer tumor cells. The IC of the benzazepine spiroindoline compound synthesized in Example 6 on breast cancer cells MCF7 50 like Figure 5 Shown is 5.28 μM.

[0167] Example 14 Flow cytometry experiment

[0168] The effect of the benzazepine spiroindoline compound synthesized in Example 6 on apoptosis of breast cancer cells (MCF7) was tested by flow cytometry for apoptosis of cultured tumor cells. The flow cytometer used was produced by Beckman coulter, and the apoptosis detection kit (Annexin V-FITC / PI) was produced by Yisheng Biology. 0μM, 1.25μM, 2.5μM, 5μM, 10μM, and 20μM of the benzazepine spiroindoline compound synthesized in Example 6 were added to treat the cells, and then immediately tested on the machine. Each was repeated three times, and a total of 18 samples were collected. The statistical graph is shown as follows Figure 6 As shown in the data, the higher the drug concentration, the more apoptotic cells there are; the flow cytometric graph is shown in Figure 7As shown, the proportion of cells in the Q2+Q3 quadrant increases with increasing drug concentration, which means that the number of apoptotic cells also increases with increasing drug concentration.

[0169] In summary, the benzazepine spiroindoline compound of the present invention can be used in the preparation of anti-tumor drugs, especially in the preparation of anti-breast cancer drugs.

[0170] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, which should be included in the scope of the claims of the present invention. The techniques, shapes, and structural parts not described in detail in the present invention are all known technologies.

Claims

1. A benzazepine spiroindoline compound, characterized in that: The general structural formula of the benzazepine spiroindoline compound is as follows: The benzazepine spiroindoline compound is selected from the following compounds:

2. A method for synthesizing a benzazepine spiroindoline compound, characterized in that: The following operations are included: S1. Compound 2, Compound 3 and Compound 4 are sequentially added to a methanol solution containing Compound 1, mixed and stirred, precipitated, and filtered to obtain a Ugi adduct, i.e., an intermediate product Compound 1, wherein Compound 1 is an aldehyde compound, Compound 2 is a tryptamine derivative, Compound 3 is an o-iodobenzoic acid derivative, and Compound 4 is an isonitrile derivative; The structural formula of the compound 1 is The structural formula of the compound 2 is The structural formula of the compound 3 is The structural formula of the compound 4 is S2. Add Ugi adduct, Pd(OAc)2, Ph3P and K2CO3 into a sealed container and stir to mix, evacuate the container and fill it with nitrogen, repeat this operation three times, then add anhydrous 1,2-dichlorobenzoate and seal the reaction, monitor by TLC, and perform wet loading and silica gel column chromatography on the obtained reaction mixture to obtain the target product benzazepine spirodihydroindole compound; The synthetic route is as follows: The structural formula of the target product benzazepine spiroindoline compound is as described in claim 1.

3. The method for synthesizing a benzazepine spiroindoline compound according to claim 2, characterized in that: In the step S1, the molar ratio of compound 1, compound 2, compound 3 and compound 4 is 1:1:1:0.

93.

4. The method for synthesizing a benzazepine spiroindoline compound according to claim 3, characterized in that: In the step S2, the molar ratio of the intermediate product compound 1, Pd(OAc)2, Ph3P, and K2CO3 is 1:0.05:0.1:2.

0.

5. The method for synthesizing a benzazepine spiroindoline compound according to claim 4, characterized in that: In the step S2, the temperature of the sealing reaction is 120° C. and the reaction time is 16 hours.

6. The method for synthesizing a benzazepine spiroindoline compound according to claim 5, characterized in that: In the step S2, the silica gel column chromatography uses ethyl acetate / n-hexane = 30% to 40% as the eluent.

7. Use of the benzazepine spiroindoline compound according to claim 1 or the benzazepine spiroindoline compound synthesized by the synthesis method according to any one of claims 2 to 6 in the preparation of an anti-tumor drug, wherein the anti-tumor drug is a drug for treating breast cancer and colon cancer.

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