CN107857766B - A kind of synthetic method and application of spiro indole compounds based on phenylalanine and polycarbonyl cyclic ketone compounds - Google Patents

Abstract

The invention discloses a synthesis method of a spiral indole compound based on phenylalanine and a polycarbonyl cyclic ketone compound. Compared with the prior art, the synthesis method is simple and convenient, the original multi-step reaction is replaced by the single-pot synthesis method, the raw materials are easy to obtain, the reaction efficiency is high, and the method is suitable for various reaction substrates; meanwhile, the obtained spiral indole compound has pharmaceutical activity and can be used in drugs which can effectively treat tumors and cancers clinically.

Description

A spiro indole compound based on phenylalanine and polycarbonyl cyclic ketone compounds Synthetic methods and applications

technical field

The invention belongs to the field of organic synthesis, in particular to a method for synthesizing spiro indole compounds based on phenylalanine and polycarbonyl cyclic ketone compounds and applications thereof.

Background technique

Spirooxindole compounds have been reported as inhibitors of P53-MDM2. These small-molecule compounds can be used as a drug for tumor therapy, directly targeting tumor cells without affecting normal cells. In the past few years, significant progress has been made in synthetic approaches to obtain spiroindole derivatives while creating a way for a chiral quaternary carbon center. For example, Overman reported an asymmetric intramolecular Heck reaction for the synthesis of spiro-pyrrole-3,3'-indole] derivatives with high correspondence selectivity; (ref. V. Caprio, B. Guyen, Y. Opoku-Boahen, J.Mann, S.M.Gowan, L.M.Kelland, M.A.Read and S.Neidle, Bio.Med.Chem.Lett., 2000, 10, 2063-2066.)

Xie and Wang reported the one-pot reaction of indole with amino acid esters and isatin. (References J. Liu, H. Sun, X. Liu, L. Ouyang, T. Kang, Y. Xie and X. Wang, Tetrahedron Lett., 2012, 53, 2336-2340.)

The methods reported above need to be modified on the indole ring in advance, and the reaction sites are also limited, so that derivatization and molecular skeleton modification cannot be carried out quickly and in large quantities, which greatly limits the research on product diversity. At present, the rapid preparation of fused-ring skeletons by multi-component synthesis and step-by-step synthesis methods has been widely used in pharmaceutical synthesis. The structure is modified to realize the construction of complex systems. But the existing synthesis technology generally needs to go through relatively complicated synthesis steps, and the price of reaction raw materials is relatively high, and there are many problems such as certain difficulty in operation. Therefore, it is necessary to provide a new synthetic method of spiroindoles.

SUMMARY OF THE INVENTION

The object of the present invention is to provide a kind of synthetic method of the spiro indole compounds based on phenylalanine and polycarbonyl cyclic ketone compounds, utilize one-pot synthesis method to replace the original multi-step reaction, the pyridyl indole compounds Addition condensation reaction with phenylalanine and polycarbonyl cyclic ketone compounds in ethanol aqueous solution can obtain spiro indole compounds in high yield.

The invention also provides the application of the spiro indole compound in testing lung cancer or gastric cancer cell proliferation experiments.

In order to achieve the above object, the technical scheme adopted in the present invention is: a kind of synthetic method of the spiro indole compound based on phenylalanine and polycarbonyl cyclic ketone compound, and the synthetic method comprises the following steps:

(1) placing pyridyl indole compound, phenylalanine and polycarbonyl cyclic ketone compound in ethanol aqueous solution reaction solvent, heating and stirring to reflux reaction; wherein, pyridyl indole compound, phenylalanine and polycarboxylate The molar ratio of the carbonyl cyclic ketone compound is 1:(0.8-1):(1-1.2);

(2) After the reaction is completed, it is cooled to room temperature, and the product is separated and purified by suction filtration and recrystallization to obtain a spiro indole compound.

In the step (1), the volume ratio of absolute ethanol and distilled water in the aqueous ethanol solution is 2:(1-1.2).

The reaction conditions of the step (1) are as follows: in an air environment, the reaction is stirred and refluxed for 2-4 hours under heating conditions.

In the step (1), the consumption ratio of the pyridyl indole compound to the reaction solvent is 1 mmol: (45-50) mL.

The specific steps of the step (2) are: after the reaction is completed, after the temperature is cooled to room temperature, the reaction product is subjected to suction filtration to obtain a solid crude product, and then the solid crude product is recrystallized with a CH ₃ OH/DMF mixed solvent, and the recrystallization is carried out. After 1-2 times, the spiro indole compound is obtained; wherein, the volume ratio of CH ₃ OH and DMF in the CH ₃ OH/DMF mixed solvent is 7:3.

其中R为-H，-CH₃，-CH₂CH₃，-CHO，-COOCH₂CH₃，-OCH₃，-CF₃，-NO₂，-NH₂，-Ph，-Cl，Br或-I中的任意一种。In step (1), the structural formula of the pyridyl indole compound is

where _R is -H, _-CH3 , -CH2CH3, -CHO, _-COOCH2CH3 , _-OCH3 , _-CF3 , _-NO2 , _-NH2 , _-Ph , -Cl, Br or _- any of I.

中的任意一种；其中，R¹为-H，-CH₃，-CH₂CH₃，-CHO，-COOCH₂CH₃，-OCH₃，-CF₃，-NO₂，-NH₂，-Ph，-Cl，Br或-I中的任意一种。In step (1), the polycarbonyl cyclic ketone compound is selected from

Any one of them; wherein, R ¹ is -H, -CH ₃ , -CH ₂ CH ₃ , -CHO, -COOCH ₂ CH ₃ , -OCH ₃ , -CF ₃ , -NO ₂ , -NH ₂ , - Any of Ph, -Cl, Br or -I.

The spiro indole compound provided by the present invention is prepared by the above method.

The invention also provides the application of the spiro indole compound in testing the proliferation of lung cancer or gastric cancer cells, that is, the product obtained by the reaction is formulated into a concentration of 0, 1.5625, 3.125, 6.25, 12.5, 25, 50, 100 μg/ml Lung cancer and gastric cancer cells were respectively tested and analyzed by cell proliferation experiments with the solution of this concentration gradient.

The reaction mechanism of the present invention: pyridyl indole first undergoes Michael addition reaction with polycarbonyl ketone, and then condenses with phenylalanine to form a spiro ring.

Beneficial effects of the present invention: compared with the prior art, the present invention adopts a one-pot synthesis method to replace the original multi-step reaction, and the pyridyl indole compound, phenylalanine and polycarbonyl cyclic ketone compound are prepared in an aqueous ethanol solution. The addition and condensation reaction can be carried out to obtain high-yield spiro indole compounds, the method is simple and convenient to operate, the raw materials are cheap and easy to obtain, the reaction solvent is environmentally friendly and pollution-free, and the synthesized spiro indole compounds can inhibit the proliferation of lung cancer or gastric cancer cells. suppress.

Description of drawings

Fig. 1 is the hydrogen nuclear magnetic resonance spectrogram of embodiment 1;

Fig. 2 is the carbon nuclear magnetic resonance spectrogram of embodiment 1;

Fig. 3 is the hydrogen nuclear magnetic resonance spectrogram of embodiment 2;

Fig. 4 is the carbon nuclear magnetic resonance spectrogram of embodiment 2;

Fig. 5 is the hydrogen nuclear magnetic resonance spectrogram of embodiment 3;

Fig. 6 is the carbon nuclear magnetic resonance spectrogram of embodiment 3;

Fig. 7 is the hydrogen nuclear magnetic resonance spectrogram of embodiment 4;

Fig. 8 is the carbon nuclear magnetic resonance spectrogram of embodiment 4;

Fig. 9 is the data graph of the concentration gradient after the preparation of the products obtained in Example 1-4 on the proliferation of lung cancer or gastric cancer cells;

Among them, IC50 (BGC-823) is the data obtained from the gastric cancer cell proliferation experiment, and IC50 (A549) is the data obtained from the lung cancer cell proliferation experiment;

Figure 10 is a graph showing the effect of the concentration gradient of the products obtained in Examples 1-4 on the activity of lung cancer or gastric cancer cells;

Wherein, the tumor chemotherapy drug cisplatin (Cis-DDP) is the positive reference control substance, DMSO is the negative reference control substance, and P1-P4 respectively correspond to the concentration gradients of the products obtained in Examples 1-4 after preparation;

Figure 11 is the reaction equation of the present invention.

Detailed ways

Specific embodiments of the present invention will be described in detail below. It should be understood that the specific embodiments described herein are only used to illustrate and explain the present invention and are not limited to these embodiments. In the embodiment, all reaction raw materials and solvents etc. are all products of Anaiji reagent.

Example 1

A kind of synthetic method of phenylalanine-based spiro indole compound, described synthetic method comprises the following steps:

(1) take by weighing 1mmol 2-vinyl pyridyl indoline, 1mmol phenylalanine and 1mmol isatin respectively, be placed in the ethanolic aqueous solution reaction solvent of 50mL, under air environment, heating and stirring to reflux for 3 hours;

(2) after the reaction is finished, after the temperature is cooled to room temperature, the reaction product is subjected to suction filtration to obtain a solid crude product, and then the solid crude product is recrystallized with a CH ₃ OH/DMF mixed solvent, and an orange-yellow solid is obtained after recrystallization 2 times Namely, the pyridyl indoline compound 1;; the yield is 50%, and the melting point is 120-124°C.

The product structural formula of the obtained pyridinoindoline compound 1 is:

1H NMR data of the product:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 10.43(s, 1H), 10.26(s, 1H), 8.28-8.15(m, 2H), 7.91-7.78(m, 1H), 7.51(d, J =7.0Hz,1H),7.24(dd,J=5.1,3.3Hz,2H),7.19-7.13(m,3H),7.13-7.05(m,2H),6.89(dtd,J=11.5,7.6,1.1 Hz,2H),6.61-6.47(m,4H),4.97(s,1H),4.03(q,J=7.1Hz,1H),3.83(d,J=8.6Hz,1H),2.98(ddd,J =18.9,13.8,6.7Hz,2H).

Carbon NMR data of the product:

¹³ C NMR(101MHz,DMSO-d ₆ )δ180.67,177.27,150.95,147.56,142.55,142.37,139.15,136.74,134.10,129.48,128.98,128.03,127.19,126.05,125.91,125.41,124.91,122.83,121.47,120.65 , 109.11, 108.98, 75.69, 65.07, 64.05, 59.77, 57.96.

Example 2

A kind of synthetic method of phenylalanine-based spiro indole compound, described synthetic method comprises the following steps:

(1) take by weighing 1mmol 2-vinyl pyridyl indoline respectively, 1mmol phenylalanine and 1mmol 4-nitro-isatin, be placed in the ethanolic aqueous solution reaction solvent of 50mL, under air environment, heating and stirring reflux reaction 3 Hour;

(2) after the reaction is finished, after the temperature is cooled to room temperature, the reaction product is subjected to suction filtration to obtain a solid crude product, and then the solid crude product is recrystallized with a CH ₃ OH/DMF mixed solvent, and a white solid is obtained after recrystallization 2 times Namely, the pyridinoindoline compound 2;; the yield is 35%, and the melting point is 80-82°C.

The product structural formula of the obtained pyridyl indoline compound 2 is:

1H NMR data of the product:

¹ H NMR (400MHz, DMSO-d ₆ )δ10.89(s,1H),10.56(s,1H),8.42(dd,J=6.0,2.3Hz,1H),8.31-8.20(m,2H), 8.11(ddd,J=8.6,4.2,1.8Hz,1H),7.78(d,J=7.6Hz,1H),7.27(d,J=7.0Hz,2H),7.24-7.14(m,3H),7.10 (dd,J＝8.6,4.8Hz,1H),6.97-6.83(m,1H),6.81-6.68(m,1H),6.60(t,J＝6.2Hz,1H),6.57-6.42(m,2H) ),4.98(d,J＝2.6Hz,1H),3.93(dd,J＝35.4,8.6Hz,2H),2.99(ddd,J＝18.0,13.5,6.4Hz,2H).

Carbon NMR data of the product:

¹³ C NMR(101MHz,DMSO-d ₆ )δ179.82,178.38,150.73,148.98,147.76,142.36,141.97,139.21,136.63,133.52,129.05,128.34,128.06,127.56,127.38,126.72,125.96,124.42,122.99,121.93 , 120.78, 109.31, 109.12, 74.95, 65.10, 63.75, 57.12.

Example 3

A kind of synthetic method of phenylalanine-based spiro indole compound, described synthetic method comprises the following steps:

(1) take by weighing 1mmol 2-vinyl pyridyl indoline respectively, 1mmol phenylalanine and 1mmol acenaphthylene quinone, be placed in the ethanolic aqueous solution reaction solvent of 50mL, under air environment, heated and stirred to reflux for 2 hours;

(2) after the reaction is finished, after the temperature is cooled to room temperature, the reaction product is subjected to suction filtration to obtain a solid crude product, and then the solid crude product is recrystallized with a CH ₃ OH/DMF mixed solvent, and an orange-yellow solid is obtained after recrystallization 2 times Namely, pyridinoindoline compound 3; the yield is 63%, and the melting point is 86-87°C.

The product structural formula of the obtained pyridyl indoline compound 3 is:

1H NMR data of the product:

¹ H NMR (400MHz, DMSO-d ₆ )δ10.23(s, 1H), 8.18(d, J=2.8Hz, 2H), 8.11-8.02(m, 1H), 7.87(d, J=8.3Hz, 1H), 7.80(d, J＝7.0Hz, 1H), 7.73(d, J＝8.0Hz, 1H), 7.69-7.51(m, 3H), 7.23(d, J＝7.1Hz, 2H), 7.20- 7.08(m,3H),7.05(t,J=7.3Hz,1H),6.69(td,J=7.7,1.1Hz,1H),6.54(d,J=7.6Hz,1H),6.47-6.35(m ,1H),6.30(d,J＝7.5Hz,1H),5.00(d,J＝1.2Hz,1H),3.98(d,J＝9.1Hz,1H),3.81(d,J＝8.1Hz,1H) ),3.01(ddd,J=18.9,13.7,6.5Hz,2H).

Carbon NMR data of the product:

¹³ C NMR(101MHz,DMSO-d ₆ )δ203.36,179.59,150.65,147.70,142.15,140.84,139.02,136.57,133.40,131.71,130.80,129.70,129.15,128.64,128.19,128.03,127.97,127.87,125.93,125.25 , 124.66, 122.92, 122.54, 121.06, 120.32, 108.94, 78.58, 65.97, 63.42, 57.43.

Example 4

A kind of synthetic method of phenylalanine-based spiro indole compound, described synthetic method comprises the following steps:

(1) respectively take by weighing 1mmol 2-vinyl pyridyl indoline, 1mmol phenylalanine and 1mmol 4-bromo-isatin, be placed in the ethanolic aqueous solution reaction solvent of 48mL, under air environment, heating and stirring reflux reaction 3 hours ;

(2) after the reaction is finished, after the temperature is cooled to room temperature, the reaction product is subjected to suction filtration to obtain a solid crude product, and then the solid crude product is recrystallized with a CH ₃ OH/DMF mixed solvent, and a white solid is obtained after the recrystallization 2 times. Pyridinoindoline compound 4; 26% yield, mp 87-89°C.

The product structural formula of the obtained pyridinoindoline compound 4 is:

1H NMR data of the product:

¹ H NMR (400MHz, DMSO-d ₆ ) δ 10.57 (s, 1H), 10.36 (s, 1H), 8.23 (dd, J=4.7, 1.5Hz, 2H), 7.81 (d, J=8.0Hz, 1H), 7.66(d, J=2.0Hz, 1H), 7.32(dd, J=8.3, 2.1Hz, 1H), 7.28-7.22(m, 2H), 7.22-7.14(m, 3H), 7.10(dd , J=8.3, 6.2Hz, 1H), 6.90 (td, J=7.6, 1.1Hz, 1H), 6.62 (d, J=7.4Hz, 1H), 6.52 (ddd, J=8.5, 5.5, 2.0Hz, 3H),4.95(dd,J=8.7,5.1Hz,1H),3.83(d,J=8.8Hz,1H),3.74(d,J=9.8Hz,1H),3.03(dd,J=13.7,8.3 Hz,1H),2.90(dd,J=13.7,4.8Hz,1H).

Carbon NMR data of the product:

¹³ C NMR(101MHz,DMSO-d ₆ )δ180.78,177.76,151.35,148.17,142.95,142.23,139.75,137.21,134.34,132.64,129.56,129.25,129.18,128.64,128.53,127.88,126.44,125.21,123.43,121.23 , 113.69, 111.50, 109.54, 75.98, 65.63, 64.57, 58.16, 56.56.

Test example

The products obtained in Examples 1-4 were added to DMSO and dissolved into a standard concentration solution of 100 μg/ml, and then diluted in half to form solutions with concentrations of 0, 1.5625, 3.125, 6.25, 12.5, 25, and 50 μg/ml, respectively, Lung cancer and gastric cancer cells were then tested for cell proliferation with this concentration gradient solution, respectively, and cell viability was measured by the cells' ability to reduce CCK-8 to blue-violet compounds. Lung cancer cell line A549 and gastric cancer cell line BGC-823 were inoculated in 96-well plates 24 h in advance, respectively, at a cell density of 3×104. After the cells were treated in groups with the above concentration gradient solution for 36-48 hours, 10 μL of CCK-8 was added to each well, and the culture was continued for 2 hours. The absorbance at 450 nm wavelength of each well was detected by a microplate reader. Through Figures 9 and 10, it can be concluded that the tumor chemotherapy drug cisplatin (Cis-DDP) is used as the positive reference control substance, and DMSO is used as the negative reference control substance. Most obviously, the product obtained in Example 4 has the most obvious inhibitory effect on the BGC-823 gastric cancer cell proliferation test.

Claims (2)

1. A spiro indole compound, The compound structural formula is:

2. The application of the spiro indole compound as claimed in claim 1 in the preparation of a medicament for the treatment of gastric cancer.

Images