Synthetic method of indole derivative and application of indole derivative in preparation of medicine for resisting children nephroblastoma
By synthesizing multi-substituted indole derivatives using electro-organic synthesis technology, the problem of limited efficacy of existing chemotherapy regimens for pediatric nephroblastoma has been solved. This approach achieves highly efficient and low-toxicity anti-tumor activity and has the potential to be developed into a novel drug.
Patent Information
- Application Number
- CN202511719918.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-21
- Publication Date
- 2026-02-10
AI Technical Summary
Existing chemotherapy regimens have limited efficacy against high-risk, recurrent, or metastatic nephroblastoma in children, and long-term chemotherapy can cause serious toxic side effects. There is a lack of novel, highly effective, and low-toxicity anti-nephroblastoma drugs in clinical practice.
By using electro-organic synthesis technology, the coupling reaction of aniline compounds and α-oxocarboxylic acids was realized to synthesize multi-substituted indole derivatives, avoiding the use of toxic strong oxidants. The inhibitory effect of the compounds on tumor cells was explored using the MTT assay.
The synthesized indole derivative significantly inhibits the growth of nephroblastoma cells and has the potential to be developed into an anti-nephroblastoma drug. Moreover, the reaction conditions are mild, the yield is high, and the environment is environmentally friendly.
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Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of biological medicine, and relates to a synthetic method of an indole derivative and application of the indole derivative in preparation of an anti-child nephroblastoma drug. BACKGROUND
[0002] An indole core structure widely exists in nature, is an advantageous skeleton of numerous natural products and drug molecules, and occupies an important position in the fields of medicinal chemistry and organic synthesis. From the most basic amino acid tryptophan to neurotransmitter serotonin and to plant hormone indole-3-acetic acid, indole derivatives play a key role in life activities (Yong Peng, et al. Journal of Medicinal Chemistry, 2025, 68, 11188; C. P. Singh, et al. European Journal of Medicinal Chemistry, 2010, 45, 3056; Peter B. Sampson, et al. Journal of Medicinal Chemistry, 2015, 58, 130.). More importantly, the structural unit is a “privileged structure” for drug design, and a large number of marketed clinical drugs contain an indole ring, for example, the non-steroidal anti-inflammatory drug indomethacin, the serotonin reuptake inhibitor sertraline, and a series of important antitumor drugs such as vinblastine and sunitinib. These molecules exhibit a wide range of biological activities including antibacterial, antiviral, anti-inflammatory, analgesic, and antitumor activities by efficiently interacting with various receptors and enzymes in the body (Xuhong Qian, et al. Bioorganic & Medicinal Chemistry, 2010, 18, 3279; Dilek Nartop, et al. Journal of Molecular Structure, 2019, 1195, 877; Ping Gong, et al. European Journal of Medicinal Chemistry, 2011, 46, 3149.). Therefore, developing novel, efficient, and green methods to construct structurally diverse polysubstituted indole derivatives not only has important theoretical significance for organic synthesis methodology, but also is a key to drug lead compound discovery and optimization, and provides an indispensable chemical entity library for the research and development of new drugs for various major human diseases (especially cancer).
[0003] Wilms' tumor is the most common renal malignancy in infants and young children, which seriously threatens the life and health of children. At present, the standard treatment regimen is a comprehensive treatment strategy centered on surgical resection combined with radiotherapy and chemotherapy (especially the ACT regimen of multiple drugs such as actinomycin D and vincristine). Although this strategy has significantly improved the overall survival rate of children, it still faces severe challenges. First, for high-risk, recurrent or metastatic children, the existing chemotherapy regimen has limited efficacy, and the tumor is prone to drug resistance, leading to treatment failure. Second, the long-term side effects of chemotherapy, such as cardiotoxicity, nephrotoxicity, neurotoxicity and secondary malignancies, cause irreversible damage to children who are in the critical period of growth and development, seriously affecting their long-term quality of life. Therefore, there is an urgent need to develop new, efficient and low-toxicity anti-Wilms' tumor chemotherapy drugs (Steffen M. Berger, et al. International Journal of Molecular Sciences, 2024, 25,2281; Hongtao Li, et al. Anti-Cancer Agents in Medicinal Chemistry, 2021, 21,1120; Qiang Wu, et al. Bioengineered, 2022, 13, 6136.). By designing and synthesizing small molecule compounds with new mechanisms of action or the ability to overcome drug resistance of existing drugs, it is expected to provide new treatment options for children who are not sensitive to standard chemotherapy or have relapsed after treatment, fill the gap in current clinical treatment, and have extremely important practical significance for improving the prognosis of Wilms' tumor in children, reducing the suffering of children and the burden on families and society.
[0004] Based on the research interest in the efficient construction method of heterocyclic compounds, we realized the coupling reaction of aniline compounds and α The coupling reaction of aniline compounds and -oxo carboxylic acids was realized by the strategy of electro-organic synthesis, and the efficient preparation of three polysubstituted indole derivatives was completed. The reaction conditions are mild, the yield is high, and the operation is simple. No toxic strong oxidizing agent is needed, which has good synthetic applicability and environmental friendliness. In addition, the synthesized polysubstituted indole derivatives show significant antitumor activity against Wilms' tumor cell lines, can significantly inhibit tumor cell proliferation, and have the potential to be further developed as anti-Wilms' tumor drugs. SUMMARY
[0005] The purpose of the present application is to provide a synthesis method of indole derivatives and its application in the preparation of anti-Wilms' tumor drugs.
[0006] The idea of the present application is to realize the coupling reaction of aniline compounds and αThe coupling reaction of α-oxocarboxylic acids avoids the use of toxic strong oxidants, and highly functionalized indole derivatives are synthesized. At the same time, the ability of the target compound to inhibit the growth of nephroblastoma cell lines is explored using the MTT assay.
[0007] The specific steps for synthesizing indole derivatives are as follows: In the unseparated electrolytic cell, aniline compounds are added. α -Oxocarboxylic acid, catalyst, electrolyte, and solvent were used. A mesh of glassy carbon was used as the anode, and a platinum sheet as the cathode. The reaction was carried out under magnetic stirring at room temperature for 1.5 hours at 8 mA. After the reaction was completed, the mixture was immediately filtered. The filtrate was extracted multiple times with ethyl acetate. The combined organic phases were then evaporated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography to obtain an indole derivative. The synthetic route is shown below:
[0008] The aniline compound in the step is 2-(4-benzyloxy-1- H -indol-1-yl)aniline and 2-(1 H One of (-indole-1-yl)aniline; The steps α -Oxocarboxylic acids include benzoylcarboxylic acid, 2-(4-bromo-2-fluorophenyl)-2-oxoacetic acid, and 2-(benzo[ b One of the following: thiophene-2-yl)-2-oxoacetic acid; The catalyst in the step is one of palladium acetate, palladium chloride, and palladium trifluoroacetate, preferably palladium trifluoroacetate; The aniline compounds in the steps and α The molar ratio of α-oxocarboxylic acids is 1:2; The molar ratio of aniline compounds to catalyst in the aforementioned step is 1:0.1; The electrolyte in the step is one of tetra-n-butylammonium iodide, tetra-n-butylammonium hexafluorophosphate and tetra-n-butylammonium acetate, preferably tetra-n-butylammonium iodide; The solvent used in the step is one of acetonitrile, methanol, and ethanol, preferably ethanol.
[0009] The indole derivatives A, B, and C in this invention are used to prepare drugs that inhibit the growth of nephroblastoma cell lines.
[0010] The principle of this invention is as follows: the amino group in aniline compounds acts as a directing group, which activates the inert CH bond through a palladium catalyst to form a seven-membered ring palladium intermediate. αThe acylation reaction of oxo carboxylic acid is realized by ligand exchange, decarboxylation and reductive elimination reaction, followed by intramolecular dehydration cyclization reaction to form the final polysubstituted indole derivative. The zero-valent palladium released after reductive elimination realizes catalytic cycle by anodic oxidation.
[0011] Compared with the prior art, the present application has the following advantages and beneficial effects: (1) The present application realizes the acylation reaction of aniline compounds and α The efficient coupling reaction of oxo carboxylic acid is realized, and the multi-functional group substituted indole derivative is synthesized under electrochemical conditions. The method has mild reaction conditions and can be completed at room temperature, and the use of toxic strong oxidants is avoided. In addition, the target product prepared by the method has high yield and good atom economy.
[0012] (2) It is found by MTT method detection and analysis that the prepared indole derivatives A, B and C can significantly inhibit the growth of nephroblastoma cell lines. BRIEF DESCRIPTION OF DRAWINGS
[0013] Figure 1 It is the nuclear magnetic resonance hydrogen spectrum of the indole derivative A in the present application; Figure 2 It is the nuclear magnetic resonance carbon spectrum of the indole derivative A in the present application; Figure 3 It is the nuclear magnetic resonance hydrogen spectrum of the indole derivative B in the present application; Figure 4 It is the nuclear magnetic resonance carbon spectrum of the indole derivative B in the present application; Figure 5 It is the nuclear magnetic resonance hydrogen spectrum of the indole derivative C in the present application; Figure 6 It is the nuclear magnetic resonance carbon spectrum of the indole derivative C in the present application. DETAILED DESCRIPTION
[0014] In order to further understand the present application, the specific embodiments of the present application are described in detail below in combination with the technical solutions and drawings, but it does not mean any limitation on the present application.
[0015] Example 1: 2-(4-benzyloxy-1 H indol-1-yl)aniline (0.2 mmol), benzoylformic acid (0.4 mmol) and tetra-n-butylammonium iodide (0.2 mmol) were weighed in turn and transferred to a 15-milliliter three-necked flask, 4 milliliters of ethanol was added, then an electrode was inserted, the anode material was RVC, the cathode material was platinum sheet, the power was turned on, the current was adjusted to 8 mA, and magnetic stirring was carried out at room temperature for 1.5 hours. After the reaction was completed, the power was turned off, and the organic phase was immediately filtered and collected. After rotary evaporation under reduced pressure, the crude product 8-(phenoxy methyl)-6-phenyl indolo[1,2-a ] quinoxaline (A), and the pure target product was obtained by column chromatography. The eluent was petroleum ether and ethyl acetate with a volume ratio of 10:1, and the separation yield was 83%. The product was a light yellow solid.
[0016] The structural characterization data of the target product obtained are as follows: 1 H NMR (400 MHz, CDCl3) δ 8.51 (d, J = 8.3 Hz, 1H), 8.18-7.98 (m, 4H),7.67-7.57 (m, 4H), 7.53 (d, J = 7.2 Hz, 2H), 7.50-7.35 (m, 6H), 6.87 (d, J = 7.8Hz, 1H), 5.30 (s, 2H); 13 C NMR (100 MHz, CDCl3) δ 156.3, 153.6, 138.1, 136.8, 136.3, 134.3,130.3, 130.1, 129.9, 128.7, 128.6, 128.1, 128.0, 128.0, 127.5, 125.4, 124.3,121.1, 114.7, 107.6, 102.8, 100.2, 70.1; The structure of the target compound is inferred from the above characterization data as follows:
[0017] Example 2: 2-(1 H indol-1-yl)aniline (0.2 mmol), 2-(4-bromo-2-fluorophenyl)-2-oxoacetic acid (0.4 mmol), and tetra-n-butylammonium iodide (0.2 mmol) were weighed in sequence and transferred to a 15-milliliter three-necked flask, 4 milliliters of ethanol was added, followed by inserting an electrode, the anode material was RVC, and the cathode material was a platinum sheet, the power was turned on, the current was adjusted to 8 mA, and magnetic stirring was performed at room temperature for 1.5 hours, the power was turned off after the reaction was completed, and the organic phase was immediately filtered and collected, and the crude product 6-(4-bromo-2-fluorophenyl)indolo[1,2- a ] quinoxaline (A), and the pure target product was obtained by column chromatography. The eluent was petroleum ether and ethyl acetate with a volume ratio of 10:1, and the separation yield was 83%. The product was a light yellow solid.
[0018] The structural characterization data of the target product is as follows: 1 H NMR (400 MHz, CDCl3) δ 8.52 (dd, J = 19.8, 8.5 Hz, 2H), 8.09 (d, J = 7.9Hz, 1H), 7.89 (d, J = 8.0 Hz, 1H), 7.67 (dd, J = 11.4, 4.3 Hz, 1H), 7.62-7.41 (m,5H), 7.27-7.19 (m, 1H), 6.79 (s, 1H); 13 C NMR (100 MHz, CDCl3) δ 162.8 (d, J = 251.0 Hz), 155.3, 135.7, 134.9(d, J = 4.0 Hz), 133.1, 131.7 (d, J = 9.0 Hz), 130.6, 130.4, 129.1, 129.1, 129.0,124.6, 124.3, 122.9 (d, J = 6.0 Hz), 122.8, 120.9, 120.6, 115.0, 114.8, 114.6,102.3. The structure of the target compound is inferred from the above characterization data as follows:
[0019] Example 3: 2-(1 H -Indol-1-yl)aniline (0.2 mmol), 2-(benzo[ b ]thiophen-2-yl)-2-oxoacetic acid (0.4 mmol) and tetra-n-butylammonium iodide (0.2 mmol) were weighed in turn and transferred to a 15-mL three-necked flask, 4 mL of ethanol was added, followed by inserting an electrode, the anode material was RVC and the cathode material was platinum sheet, the power was turned on, the current was adjusted to 8 mA, and magnetic stirring was carried out at room temperature for 1.5 hours. After the reaction was completed, the power was turned off, and the organic phase was immediately filtered and collected. After rotary evaporation under reduced pressure, the crude product 6-(benzo[ b ]thiophen-2-yl)indolo[1,2- aQuinoxaline (C), and the pure target product was finally separated by column chromatography. The eluent was petroleum ether and ethyl acetate with a volume ratio of 10:1, and the separation yield was 78%. The product was a yellow solid.
[0020] The structural characterization data of the obtained target product are as follows: 1 H NMR (400 MHz, CDCl3) δ 8.42 (dd, J = 8.2, 6.1 Hz, 2H), 8.24 (s, 1H),8.05 (dd, J = 7.9, 1.3 Hz, 1H), 7.93 (ddd, J = 10.0, 8.8, 6.0 Hz, 3H), 7.62-7.50(m, 3H), 7.47-7.39 (m, 4H); 13 C NMR (100 MHz, CDCl3) δ 148.9, 142.2, 140.7, 140.0, 135.5, 132.9,130.4, 129.9, 129.2, 128.9, 127.6, 125.8, 125.4, 124.6, 124.6, 124.5, 124.2,122.8, 122.7, 122.3, 114.5, 101.5 The structure of the target compound is inferred from the above characterization data as follows:
[0021] Application Example 1: The inhibitory effects of the indole derivatives (A, B, and C) prepared in Examples 1, 2, and 3 on the proliferation of children's nephroblastoma cells (G-401) were detected: The anti-tumor activity (measured by IC 50 ) of the target compounds was detected by the MTT method, and the specific operation steps are as follows: First, the target cells (G-401) were cultured, and cells in the logarithmic growth phase were selected to prepare a cell suspension with a concentration of about 50,000 cells per milliliter. The culture medium used for culture was DMEM medium with the addition of fetal bovine serum. The prepared cell suspension was inoculated into a 96-well plate, and the inoculation amount of each well was 100 microliters. Subsequently, the 96-well plate was placed in a culture box at 37 degrees Celsius with 5% carbon dioxide for a period of time. During the culture process, close observation was required, and when the cell density reached about 70%, the drug addition operation could be performed.
[0022] The drugs A, B and C to be tested were added to the corresponding wells, and 4 replicates were set for each drug. The concentration range of the test compounds was set between 10 -6 and 10 -4 M. At the same time, a blank control group (Control) was set, which did not add drugs. After the addition of drugs was completed, the 96-well plate was placed back into the incubator for continued incubation for 48 hours.
[0023] After the incubation time was over, the culture solution in each well was removed, and each well was washed once with DMEM culture solution. Then, MTT was used for staining, 20 microliters of MTT solution with a concentration of 5 mg / mL was added to each well, and the staining was continued for 4 hours. After the staining was completed, the liquid in each well was aspirated, and 100 microliters of chromatographically pure DMSO was added to each well. After the addition of DMSO, the 96-well plate was placed on a shaker for 10 minutes.
[0024] Finally, the OD value of each well in the 96-well plate was detected using a microplate reader, and the detection wavelength was set to 490 nanometers. The calculation of the results was performed by SPSS software, so as to obtain the half lethal amount (IC 50 ) of each compound on the selected tumor cells. The inhibition results of the target compounds on the childhood nephroblastoma cells are shown in Table 1: Table 1: IC 50 of target compounds on childhood nephroblastoma cells (G-401) ; The experimental results show that the target indole derivatives have obvious inhibitory effect on the growth of childhood nephroblastoma cells (G-401), and the inhibitory effect of compound B is the best. The IC 50 is 11.9 ± 0.8 μM, which is expected to be developed as a drug for inhibiting the growth of nephroblastoma cell lines.
Claims
1. A method for synthesizing an indole derivative and its application in the preparation of drugs for treating pediatric nephroblastoma, characterized in that, The specific synthesis steps are as follows: In the unseparated electrolytic cell, aniline compounds are added. α -Oxocarboxylic acid, catalyst, electrolyte, and solvent were used, with reticulated glassy carbon as the anode and a platinum sheet as the cathode. The reaction was carried out under magnetic stirring at room temperature for 1.5 hours at a current of 8 mA. After the reaction was completed, the mixture was immediately filtered, and the filtrate was extracted multiple times with ethyl acetate. The combined organic phases were then evaporated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography to obtain an indole derivative, the chemical structure of which is as follows: ； The aniline compound in the step is 2-(4-benzyloxy-1- H -indol-1-yl)aniline and 2-(1 H One of (-indole-1-yl)aniline; The steps α -Oxocarboxylic acids include benzoylcarboxylic acid, 2-(4-bromo-2-fluorophenyl)-2-oxoacetic acid, and 2-(benzo[ b One of the following: thiophene-2-yl)-2-oxoacetic acid; The catalyst in the step is one of palladium acetate, palladium chloride, and palladium trifluoroacetate. The aniline compounds in the steps and α The molar ratio of α-oxocarboxylic acids is 1:2; The molar ratio of aniline compounds to catalyst in the aforementioned step is 1:0.1; The electrolyte used in the step is one of tetra-n-butylammonium iodide, tetra-n-butylammonium hexafluorophosphate, and tetra-n-butylammonium acetate. The solvent used in the step is one of acetonitrile, methanol, and ethanol.
2. Indole derivatives A, B, and C are used in the preparation of drugs that inhibit the growth of nephroblastoma cell lines.