Selective Synthesis of Arylylsulfonated Tetrahydropyrido[1,2-a]indole

Abstract

This invention discloses a green, efficient, and selective method for synthesizing arylsulfonated tetrahydropyrido[1,2-a]indole, relating to the fields of organic synthesis technology and antitumor activity evaluation. Using DABCO·(SO2)2 as the SO2 source and acetone or 1,2-dichloroethane as the solvent, this invention achieves a tandem cyclization reaction of N-alkenyl-substituted indole initiated by aryl sulfone radicals under visible light conditions, selectively yielding arylsulfonated tetrahydropyrido[1,2-a]indole products with different reaction sites. Preliminary antitumor activity evaluation of the obtained arylsulfonated tetrahydropyrido[1,2-a]indole products was then conducted, revealing that product 3wd exhibited significant inhibitory activity against mouse skin melanoma cells B16F10. The synthetic method of this invention is green, efficient, and selective, and product 3wd exhibits significant inhibitory activity against B16F10 cells.

Description

Technical Field

[0001] This invention relates to the fields of organic synthesis technology and antitumor activity evaluation. Background Technology

[0002] Since aryl sulfonyl fragments [-SO2Ar] are widely present in natural product molecules and play an important role in pharmaceuticals, pesticides, materials, and synthetic chemistry, introducing aryl sulfonyl fragments into organic molecules is of great significance. Compared with direct aryl sulfonation methods, aryl sulfonation using a combination of aryl diazonium salts and sulfur dioxide insertion strategies is more flexible and efficient.

[0003] The tetrahydropyrido[1,2-a]indole skeleton exists in many bioactive compounds and drug molecules. Currently, no method has been found to introduce [-SO2Ar] into the pyrrolo[1,2-a]indole skeleton under visible light to form arylsulfonated pyrrolo[1,2-a]indole. Therefore, developing a green and efficient method for synthesizing arylsulfonated tetrahydropyrido[1,2-a]indole is of great significance. Summary of the Invention

[0004] The purpose of this invention is to provide a green, efficient, and selective method for synthesizing arylsulfonated tetrahydropyrido[1,2-a]indole. Using DABCO·(SO₂)₂ as the SO₂ source and acetone or 1,2-dichloroethane as the solvent, the aryl sulfonation reaction of N-alkenyl-substituted indole is achieved under visible light conditions, selectively yielding arylsulfonated tetrahydropyrido[1,2-a]indole products with different reaction sites. This synthetic method is green, efficient, and selective, with some products exhibiting significant inhibitory activity against B16F10 cells.

[0005] To achieve the above objectives, the technical solution of the present invention is as follows:

[0006] In a first aspect, the present invention provides a selective method for synthesizing arylsulfonated tetrahydropyrido[1,2-a]indole, wherein an N-alkenyl-substituted indole of formula (1) is reacted with DABCO·(SO2)2 of formula (2) and an aryl diazonium salt as reactants in a reaction solvent to selectively obtain arylsulfonated tetrahydropyrido[1,2-a]indole products as shown in formulas (3) and (4);

[0007]

[0008] Where: n = 1 or 2; R 1 For H, Cl, or Me; R 2 For Ac;R 3 and R 4 For H; R 5 For Me or OMe;

[0009] The molar ratio of the N-alkenyl-substituted indole, DABCO·(SO2)2, and aryl diazonium salt is 0.2:0.4:0.6;

[0010] When obtaining product 3 as shown in formula (3), the reaction solvent is acetone;

[0011] When obtaining product 4 as shown in formula (4), the additive is formic acid and the reaction solvent is 1,2-dichloroethane.

[0012] Furthermore, the arylsulfonated tetrahydropyrido[1,2-a]indole shown in formula (3) is synthesized by the following steps:

[0013] In an argon atmosphere, 0.2 mmol of N-alkenyl-substituted indole, 0.4 mmol of DABCO·(SO2)2 and 0.6 mmol of aryl diazonium salt were added sequentially to a 25 mL reaction tube, followed by 2 mL of acetone. The mixture was stirred at 10 °C under 450 nm blue light for 24 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridino[1,2-a]indole of formula (3) was obtained by column chromatography.

[0014] Furthermore, the arylsulfonated tetrahydropyrido[1,2-a]indole shown in formula (4) is synthesized by the following steps:

[0015] In an argon atmosphere, 0.2 mmol of N-alkenyl-substituted indole, 0.4 mmol of DABCO·(SO2)2, 0.6 mmol of aryl diazonium salt and 0.4 mmol of formic acid were added sequentially to a 25 mL reaction tube, followed by 2 mL of 1,2-dichloroethane. The mixture was stirred at 10 °C under 450 nm blue light for 24 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridino[1,2-a]indole of formula (4) was obtained by column chromatography.

[0016] Further, the N-alkenyl-substituted indole is 1-(1-(4-penten-1-yl)-1H-indol-3-yl) acetone, 1-(4-chloro-1-(4-penten-1-yl)-1H-indol-3-yl) acetone, 1-(7-methyl-1-(4-penten-1-yl)-1H-indol-3-yl) acetone, 1-[1-(4-methylpent-3-en-1-yl)-1H-indol-3-yl] acetone, 1-[4-chloro-1-(4-methylpent-3-en-1-yl)-1H-indol-3-yl] acetone, 1-[7-methyl-1-(4-methylpent-3-en-1-yl)-1H-indol-3-yl] acetone, Vemurafenib-1w, and Vemurafenib-1v. The aryl diazonium salt is p-methylbenzenetetrafluoroborate and o-methoxybenzenetetrafluoroborate.

[0017] In a second aspect, the present invention also provides arylsulfonated tetrahydropyrido[1,2-a]indole, which is prepared by any of the synthetic methods described above.

[0018] Thirdly, the present invention also provides arylsulfonated tetrahydropyrido[1,2-a]indole 3wd, having the following structural formula:

[0019]

[0020] Furthermore, the synthesis method of the arylsulfonated tetrahydropyrido[1,2-a]indole 3wd includes the following steps:

[0021] In an argon atmosphere, 0.2 mmol of 1-(1-(4-penten-1-yl)-1H-indol-3-yl) ethyl ketone, 0.4 mmol of DABCO·(SO2)2, and 0.6 mmol of o-methoxybenzene tetrafluoroborate were added sequentially to a 25 mL reaction tube, followed by 2 mL of acetone. The mixture was stirred at 10 °C under 450 nm blue light for 24 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridino[1,2-a]indole 3wd was obtained by column chromatography.

[0022] Fourthly, the present invention also provides the application of the above-mentioned arylsulfonated tetrahydropyrido[1,2-a]indole 3wd in the preparation of antitumor active drugs.

[0023] Compared with the prior art, the technical effects of the present invention are as follows:

[0024] This invention provides a green, efficient, and selective method for the synthesis of arylsulfonated tetrahydropyrido[1,2-a]indole. By employing N-alkenyl-substituted indole and using DABCO·(SO2)2 as the SO2 source, arylsulfonated tetrahydropyrido[1,2-a]indole 3 and 4 can be selectively synthesized under visible light conditions. This type of reaction is green and efficient, and some products (such as 3wd) show potential for drug development in terms of antitumor activity. Attached Figure Description

[0025] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in this invention. For those skilled in the art, other drawings can be obtained based on these drawings.

[0026] Figure 1 and Figure 2 The proton and carbon spectra of compound 3wa provided in Example 1 of this invention are shown.

[0027] Figure 3 and Figure 4 The proton and carbon spectra of compound 3wb provided in Example 2 of this invention are shown.

[0028] Figure 5 and Figure 6 The proton and carbon spectra of compound 3wc provided in Example 3 of this invention are shown.

[0029] Figure 7 and Figure 8 The proton and carbon spectra of compound 3wd provided in Example 4 of this invention are shown.

[0030] Figure 9 and Figure 10 The proton and carbon spectra of compound 4va provided in Example 5 of this invention are shown.

[0031] Figure 11 and Figure 12 The proton and carbon spectra of compound 4vb provided in Example 6 of this invention.

[0032] Figure 13 and Figure 14 The proton and carbon spectra of compound 4vc provided in Example 7 of this invention.

[0033] Figure 15 and Figure 16 The proton and carbon spectra of compound 4vd provided in Example 8 of this invention are shown.

[0034] Figures 17-19 The proton, carbon, and fluorine spectra of the compound Vemurafenib-3w provided in Example 9 of this invention.

[0035] Figures 20-22 The proton, carbon, and fluorine spectra of the compound Vemurafenib-4v provided in Example 10 of this invention are shown.

[0036] Figure 23 The fluorescence spectra of the compounds 3wa~3wd, Vemurafenib-3w, 4va~4vd and Vemurafenib-4v of this invention are shown.

[0037] Figure 24 This is the fluorescence spectrum of compound 3wb of the present invention.

[0038] Figure 25 This is a linear graph of compound 3wb of the present invention.

[0039] Figure 26 The compounds 3wd, 4vd, Vemurafenib-3w, and Vemurafenib-4v of this invention were imaged in B16F10 cells.

[0040] Figure 27 The effects of the compounds 3wd, 4vd, Vemurafenib-3w, and Vemurafenib-4v of this invention on the proliferation activity of B16F10 cells were detected using the MTT assay. Detailed Implementation

[0041] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0042] Example 1

[0043] In an argon atmosphere, 0.2 mmol of 1-(1-(4-penten-1-yl)-1H-indol-3-yl) acetone (1w), 0.4 mmol of DABCO·(SO2)2, and 0.6 mmol of p-methylbenzenetetrafluoroborate (2) were added sequentially to a 25 mL reaction tube, followed by 2 mL of acetone. The mixture was stirred at 10 °C under 450 nm blue light for 24 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridino[1,2-a]indole (3wa) was obtained by column chromatography. 59.5 mg of the yellow oil was obtained, with a yield of 78%. The proton and carbon spectra of compound 3wa are shown below. Figure 1 and 2As shown. The reaction equation is as follows:

[0044]

[0045] Example 1 investigated the applicability of the substrate when the aryl diazonium salt has an electron-donating methyl group at the C4 position. The results of Example 1 showed that this substrate is also suitable for this reaction to give arylsulfonylated tetrahydropyrido[1,2-a]indole (3wa).

[0046] Example 2

[0047] In an argon atmosphere, 0.2 mmol of 1-(4-chloro-1-(4-penten-1-yl)-1H-indol-3-yl) ethyl ketone (1b), 0.4 mmol of DABCO·(SO2)2, and 0.6 mmol of p-methylbenzene tetrafluoroborate (2) were added sequentially to a 25 mL reaction tube, followed by 2 mL of acetone. The mixture was stirred at 10 °C under 450 nm blue light for 24 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridino[1,2-a]indole (3wb) was obtained by column chromatography. 68.2 mg of a yellow solid was obtained, with a yield of 82%. The proton and carbon spectra of compound 3wb are shown below. Figure 3 and 4 As shown. The reaction equation is as follows:

[0048]

[0049] Example 2 investigated the applicability of the substrate when the C4 position of the N-alkenyl-substituted indole contains an electron-withdrawing chlorine group. The results of Example 2 showed that this substrate is also suitable for this reaction to give arylsulfonylated tetrahydropyridino[1,2-a]indole (3wb).

[0050] Example 3

[0051] In an argon atmosphere, 0.2 mmol of 1-(7-methyl-1-(4-penten-1-yl)-1H-indol-3-yl) ethyl ketone (1c), 0.4 mmol of DABCO·(SO2)2, and 0.6 mmol of p-methylbenzenetetrafluoroborate (2) were added sequentially to a 25 mL reaction tube, followed by 2 mL of acetone. The mixture was stirred at 10 °C under 450 nm blue light for 24 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridino[1,2-a]indole (3wc) was obtained by column chromatography. 64.0 mg of white solid was obtained, with a yield of 81%. The proton and carbon spectra of compound 3wc are shown below. Figure 5 and 6As shown. The reaction equation is as follows:

[0052]

[0053] Example 3 investigated the applicability of the substrate when the C7 position of the N-alkenyl-substituted indole contains an electron-donating methyl group. The results showed that this substrate is also suitable for this reaction to yield arylsulfonylated tetrahydropyridino[1,2-a]indole (3wc).

[0054] Example 4

[0055] In an argon atmosphere, 0.2 mmol of 1-(1-(4-penten-1-yl)-1H-indol-3-yl) ethyl ketone (1w), 0.4 mmol of DABCO·(SO2)2, and 0.6 mmol of o-methoxybenzenetetrafluoroborate (2d) were added sequentially to a 25 mL reaction tube, followed by 2 mL of acetone. The mixture was stirred at 10 °C under 450 nm blue light for 24 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridino[1,2-a]indole (3wd) was obtained by column chromatography. 65.3 mg of white solid was obtained, with a yield of 82%. The 1H and 1C spectra of compound 3wd are shown below. Figure 7 and 8 As shown. The reaction equation is as follows:

[0056]

[0057] Example 4 primarily investigated the applicability of the substrate when the C2 position of the aryl diazonium salt contains an electron-withdrawing methoxy group. The results of Example 4 show that this substrate is also suitable for this reaction to yield arylsulfonylated tetrahydropyridino[1,2-a]indole (3wd).

[0058] Example 5

[0059] In an argon atmosphere, 0.2 mmol of 1-[1-(4-methylpent-3-en-1-yl)-1H-indol-3-yl]acetone (1v), 0.4 mmol of DABCO·(SO2)2, 0.4 mmol of formic acid, and 0.6 mmol of p-methylbenzenetetrafluoroborate (2) were added sequentially to a 25 mL reaction tube, followed by 2 mL of acetone. The mixture was stirred at 10 °C under 450 nm blue light for 24 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridino[1,2-a]indole (4va) was obtained by column chromatography. 66.6 mg of a yellow solid was obtained, with a yield of 73%. The proton and carbon spectra of compound 4va are shown below. Figure 9 and10 As shown. The reaction equation is as follows:

[0060]

[0061] Example 5 primarily investigated the applicability of the substrate as an N-4-methylpentenyl-substituted indole. The results showed that this substrate is also suitable for this reaction to yield arylsulfonylated tetrahydropyridino[1,2-a]indole (4va).

[0062] Example 6

[0063] In an argon atmosphere, 0.2 mmol of 1-[4-chloro-1-(4-methylpent-3-en-1-yl)-1H-indol-3-yl]acetone (1b'), 0.4 mmol of DABCO·(SO2)2, 0.4 mmol of formic acid, and 0.6 mmol of p-methylbenzenetetrafluoroborate (2) were added sequentially to a 25 mL reaction tube, followed by 2 mL of 1,2-dichloroethane. The mixture was stirred for 24 h under 450 nm blue light at 10 °C. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyrido[1,2-a]indole (4vb) was obtained by column chromatography. 30.9 mg of the yellow oil was obtained, with a yield of 36%. The proton and carbon spectra of compound 4vb are shown below. Figure 11 and 12 As shown. The reaction equation is as follows:

[0064]

[0065] Example 6 investigated the applicability of the substrate when the C4 position of the N-4-methylpentenyl-substituted indole contains an electron-withdrawing chlorine group. The results of Example 6 showed that the substrate is also suitable for this reaction to give arylsulfonylated tetrahydropyrido[1,2-a]indole (4vb).

[0066] Example 7

[0067] In an argon atmosphere, 0.2 mmol of 1-[7-methyl-1-(4-methylpent-3-en-1-yl)-1H-indol-3-yl]acetone (1c'), 0.4 mmol of DABCO·(SO2)2, 0.4 mmol of formic acid, and 0.6 mmol of p-methylbenzenetetrafluoroborate (2) were added sequentially to a 25 mL reaction tube, followed by 2 mL of 1,2-dichloroethane. The mixture was stirred for 24 h under 450 nm blue light at 10 °C. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyrido[1,2-a]indole (4vc) was obtained by column chromatography. 41.0 mg of white solid was obtained, with a yield of 50%. The proton and carbon spectra of compound 4vc are shown below. Figure 13 and 14 As shown. The reaction equation is as follows:

[0068]

[0069] Example 7 investigated the applicability of the substrate when the C7 position of the N-4-methylpentenyl-substituted indole contains an electron-donating methyl group. The results of Example 7 showed that the substrate is also suitable for this reaction to yield arylsulfonylated tetrahydropyridino[1,2-a]indole (4vc).

[0070] Example 8

[0071] In an argon atmosphere, 0.2 mmol of 1-[1-(4-methylpent-3-en-1-yl)-1H-indole-3-yl ethyl ketone (1v), 0.4 mmol of DABCO·(SO2)2, 0.4 mmol of formic acid, and 0.6 mmol of o-methoxybenzenetetrafluoroborate (2d) were added sequentially to a 25 mL reaction tube, followed by 2 mL of 1,2-dichloroethane. The mixture was stirred for 24 h under 450 nm blue light at 10 °C. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyrido[1,2-a]indole (4vd) was obtained by column chromatography. 52.6 mg of the yellow oil was obtained, with a yield of 64%. The 1H and 1C spectra of compound 4vd are shown below. Figure 15 and 16 As shown. The reaction equation is as follows:

[0072]

[0073] Example 8 primarily investigated the applicability of aryl diazonium salts with an electron-withdrawing methoxy group at the C2 position. The results of Example 8 indicate that this substrate is also suitable for this reaction to yield arylsulfonylated tetrahydropyridino[1,2-a]indole (4vd).

[0074] Example 9

[0075] In an argon atmosphere, 0.2 mmol of Vemurafenib-1w, 0.4 mmol of DABCO·(SO2)2, and 0.6 mmol of p-methylbenzenetetrafluoroborate (2) were added sequentially to a 25 mL reaction tube, followed by 2 mL of 1,2-dichloroethane. The mixture was stirred for 24 h under 450 nm blue light at 10 °C. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridinium[3,2-b]indene (Vemurafenib-3w) was obtained by column chromatography. 123.4 mg of white solid was obtained, with a yield of 87%. The proton, carbon, and fluorine spectra of compound Vemurafenib-3w are shown below. Figure 17 , 18 As shown in Figure 19. The reaction equation is as follows:

[0076]

[0077] Example 9 primarily investigated the suitability of the substrate for the N1-position substitution of the drug Vemurafenib with an enpentyl group. The results of this example indicate that the substrate is also suitable for this reaction to yield arylsulfonated tetrahydropyridinium[3,2-b]indene (Vemurafenib-3w).

[0078] Example 10

[0079] In an argon atmosphere, 0.2 mmol of Vemurafenib-1v, 0.4 mmol of DABCO·(SO2)2, 0.4 mmol of formic acid, and 0.6 mmol of p-methylbenzenetetrafluoroborate (2) were added sequentially to a 25 mL reaction tube, followed by 2 mL of 1,2-dichloroethane. The mixture was stirred for 24 h under 450 nm blue light at 10 °C. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridinium[3,2-b]indene (Vemurafenib-4v) was obtained by column chromatography. 79.8 mg of white solid was obtained, with a yield of 55%. The proton, carbon, and fluorine spectra of the compound Vemurafenib-4v are shown below. Figure 20 , 21 As shown in Figure 22, the reaction equation is as follows:

[0080]

[0081] Example 10 primarily investigated the suitability of the substrate for the N1-position substitution of the drug Vemurafenib with a 4-methylpentenyl group. The results of this example indicate that the substrate is also suitable for this reaction to yield arylsulfonated tetrahydropyridinium[3,2-b]indene (Vemurafenib-4v).

[0082] The fluorescence properties and antitumor activity of the technical solution described in this invention will be further verified below:

[0083] Fluorescence spectral analysis of compounds 3wa~3wd, Vemurafenib-3w, 4va~4vd, and Vemurafenib-4v, such as Figure 23 As shown: Experiments were conducted using an F-7100 fluorescence spectrophotometer. 0.5 mM solutions of 1,4-dioxane, dichloromethane (DCM), dimethyl sulfoxide (DMSO), ethanol (EtOH), phosphate buffered saline (PBS), and toluene were added to 1 cm quartz cuvettes, and excited in the range of 300–420 nm. The optimal result is shown in the fluorescence curve at 300 nm, as illustrated below. Figure 23 As shown.

[0084] like Figure 24 and 25 As shown, with the increase of the proportion of DMSO in the solvent (i.e., the system polarity increases), the fluorescence intensity of compound 3wb shows a significant increasing trend, which can be used as a polar probe (y=23.7406x-21.5647, R). 2 =0.9974).

[0085] Imaging of compounds 3wd, 4vd, Vemurafenib-3w, and Vemurafenib-4v in B16F10 cells: Logarithmic growth phase B16F10 cells were washed with PBS, digested with trypsin, and then dispersed by pipetting after adding serum-containing complete medium. After centrifugation, the cells were resuspended in complete medium, counted, and the cell density was adjusted before seeding into confocal microscopy dishes. Cells were cultured at 37°C and 5% CO2 for 24 h to allow for stable cell adhesion. The medium was then discarded, and serum-free medium containing an appropriate concentration of the probe or its derivative was added for further incubation. After incubation, the cells were slowly washed several times with PBS to remove uninfiltrated free compounds. Images were then acquired and analyzed using a laser confocal microscope. 3wd, 4vd, Vemurafenib-3w, and Vemurafenib-4v produced strong and clear fluorescence signals within the cells, mainly distributed in the cytoplasm. The cells showed clear outlines and intact morphology, indicating good cell permeability and intracellular imaging capabilities. Figure 26 As shown.

[0086] To systematically evaluate the inhibitory activity of 3wd, 4vd, Vemurafenib-3w, and Vemurafenib-4v on melanoma B16F10 cells, this invention selected the melanoma treatment drugs Vemurafenib and temozolomide as positive controls, and conducted relevant antitumor activity assessments using the MTT assay.

[0087] Logarithmic growth phase B16F10 cells were washed with PBS, digested with trypsin, and then dispersed by pipetting after adding serum-containing complete medium. After centrifugation, the cells were resuspended in complete medium, counted, and the cell density was adjusted to approximately 3000 cells / 100 μL. Cells were seeded at 100 μL / well in 96-well plates and cultured at 37℃ and 5% CO2 for 24 h. After cell attachment, medium containing different concentrations of the probe was added for treatment, with final concentrations of 0, 6.25, 12.5, 25, 50, and 100 μM, and each group was replicated. After 24 h of incubation, 20 μL of MTT solution (5 mg / mL) was added to each well for another 4 h of incubation. After discarding the culture medium, 150 μL of DMSO was added to dissolve formazan crystals, and the OD value was measured at 570 nm. With the control group cell viability as 100%, cell survival rate was calculated to evaluate the cytotoxicity and proliferation inhibition of the probe. MTT results showed that only compound 3wd (IC50) showed minimal cytotoxicity. 50 =19.38) and the positive drug Vemurafenib (IC 50 =8.673) showed a significant inhibitory effect on B16F10 cells, and the inhibitory effect of 3wd was close to that of Vemurafenib, while compounds 4vd, Vemurafenib-3w, Vemurafenib-4v and the positive control drug Temozolomide did not show significant inhibitory activity against B16F10 cells. Figure 27 As shown.

[0088] The above description is merely a preferred embodiment of the present invention and is not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention are included within the scope of protection of the present invention.

Claims

1. A selective method for synthesizing arylsulfonated tetrahydropyrido[1,2-a]indole, characterized in that, Using N-alkenyl-substituted indole as shown in formula (1) and DABCO·(SO2)2 and aryl diazonium salt as shown in formula (2) as reaction raw materials in a reaction solvent, arylsulfonated tetrahydropyrido[1,2-a]indole products as shown in formulas (3) and (4) are selectively obtained. ； Where: n = 1 or 2; R 1 For H, Cl, or Me; R 2 For Ac;R 3 and R 4 For H; R 5 For or Me or OMe; The molar ratio of the N-alkenyl-substituted indole, DABCO·(SO2)2, and aryl diazonium salt is 0.2:0.4:0.6; When obtaining product 3 as shown in formula (3), the reaction solvent is acetone; When obtaining product 4 as shown in formula (4), the additive is formic acid and the reaction solvent is 1,2-dichloroethane.

2. The selective arylsulfonated tetrahydropyrido[1,2-a]indole synthesis method according to claim 1, characterized in that, The arylsulfonated tetrahydropyrido[1,2-a]indole shown in formula (3) is synthesized by the following steps: In an argon atmosphere, 0.2 mmol of N-alkenyl-substituted indole, 0.4 mmol of DABCO·(SO2)2 and 0.6 mmol of aryl diazonium salt were added sequentially to a 25 mL reaction tube, followed by 2 mL of acetone. The mixture was stirred at 10 °C under 450 nm blue light for 24 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridino[1,2-a]indole of formula (3) was obtained by column chromatography.

3. The selective arylsulfonated tetrahydropyrido[1,2-a]indole synthesis method according to claim 1, characterized in that, The arylsulfonated tetrahydropyrido[1,2-a]indole shown in formula (4) is synthesized by the following steps: In an argon atmosphere, 0.2 mmol of N-alkenyl-substituted indole, 0.4 mmol of DABCO·(SO2)2, 0.6 mmol of aryl diazonium salt and 0.4 mmol of formic acid were added sequentially to a 25 mL reaction tube, followed by 2 mL of 1,2-dichloroethane. The mixture was stirred at 10 °C under 450 nm blue light for 24 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridino[1,2-a]indole of formula (4) was obtained by column chromatography.

4. The selective arylsulfonated tetrahydropyrido[1,2-a]indole synthesis method according to claim 1, characterized in that, The N-alkenyl-substituted indole is selected from 1-(1-(4-penten-1-yl)-1H-indole-3-yl) acetone, 1-(4-chloro-1-(4-penten-1-yl)-1H-indole-3-yl) acetone, 1-(7-methyl-1-(4-penten-1-yl)-1H-indole-3-yl) acetone, 1-[1-(4-methylpent-3-en-1-yl)-1H-indole-3-yl) acetone, etc. One of 1-[4-chloro-1-(4-methylpent-3-en-1-yl)-1H-indol-3-yl] ethyl ketone, 1-[7-methyl-1-(4-methylpent-3-en-1-yl)-1H-indol-3-yl] ethyl ketone, vemurafenib-1w, and vemurafenib-1v; wherein the aryl diazonium salt is one of p-methylbenzenetetrafluoroborate and o-methoxybenzenetetrafluoroborate.

5. Arylsulfonated tetrahydropyrido[1,2-a]indole, characterized in that, It was prepared by the synthesis method described in any one of claims 1-3.

6. Arylsulfonyl tetrahydropyrido[1,2-a]indole 3wd, characterized in that, It has the following structural formula: 。 7. The arylsulfonated tetrahydropyrido[1,2-a]indole 3wd as described in claim 6, characterized in that, The synthesis method includes the following steps: In an argon atmosphere, 0.2 mmol of 1-(1-(4-penten-1-yl)-1H-indol-3-yl) ethyl ketone, 0.4 mmol of DABCO·(SO2)2, and 0.6 mmol of o-methoxybenzene tetrafluoroborate were added sequentially to a 25 mL reaction tube, followed by 2 mL of acetone. The mixture was stirred at 10 °C under 450 nm blue light for 24 h. The reaction was monitored by thin-layer chromatography. After the reaction was completed, the mixture was quenched with water, extracted three times with ethyl acetate, washed once with salt, dried with anhydrous sodium sulfate, and the solvent was removed by vacuum distillation. The arylsulfonated tetrahydropyridino[1,2-a]indole 3wd was obtained by column chromatography.

8. The use of the arylsulfonated tetrahydropyrido[1,2-a]indole 3wd as described in claim 6 or 7 in the preparation of antitumor active drugs.

Images