Trisubstituted anisoles containing an n-propylol indole structure, and methods of making and using the same

Abstract

The present application relates to the preparation method and application of the tri-substituted anisoles containing N-propyl alcohol group indole structure, and the compounds have the structural formula shown in general formula (I). The present application relates to the compounds of general formula (I) having the effect of strongly inhibiting L858R / T790M / C797S and Del19 / T790M / C797S drug-resistant mutant EGFR kinase, and further relates to the use of the compounds and pharmaceutically acceptable salts thereof in the preparation of drugs for treating and / or preventing diseases caused by drug-resistant mutations of EGFR kinase, especially in the preparation of drugs for treating and / or preventing cancer. The present application also provides the preparation method of the compounds, and the pharmaceutical compositions and uses containing them. Especially for preparing drugs for treating and / or preventing lung cancer, having good application prospect of anti-tumor drug development. (I)

Description

Technical Field

[0001] This invention belongs to the field of compound preparation technology, specifically relating to trisubstituted anisole compounds containing N-propanol indole structures, their preparation methods, and applications. Background Technology

[0002] Lung cancer, a major disease burden threatening global public health, has long been the leading cause of cancer incidence and mortality. Non-small cell lung cancer (NSCLC) accounts for approximately 80%–85% of all lung cancers, and its clinical treatment strategies have entered the era of precision medicine guided by molecular subtyping. Among the driver genes of NSCLC, activating mutations of epidermal growth factor receptor (EGFR) have significant clinical implications—approximately 40%–55% of Asian patients carry such mutations, with exon 19 deletion (Del19) and exon 21 L858R point mutations accounting for over 90% of sensitive mutations. After EGFR activation, it drives tumor proliferation, survival, and metastasis through downstream signaling pathways (such as RAS / RAF / MEK / ERK, PI3K / AKT / mTOR). Therefore, targeted inhibition of EGFR has become a core direction for NSCLC treatment.

[0003] Since the advent of gefitinib, the first EGFR tyrosine kinase inhibitor (EGFR-TKI), the development of this class of drugs has iterated to the third generation. While first-generation reversible inhibitors showed significant efficacy, most patients developed resistance due to the T790M mutation after 9–14 months of treatment. Second-generation covalent inhibitors, although able to overcome T790M, caused significant toxicity due to their strong inhibition of wild-type EGFR. Third-generation drugs are more selective, effective against the T790M mutation, and have reduced toxicity, becoming the standard treatment, with osimertinib being the most representative. However, after 18–24 months of use with third-generation drugs, approximately 7%–25% of patients develop secondary C797S mutations, forming Del19 / T790M / C797S or L858R / T790M / C797S triple mutations, leading to the ineffectiveness of all existing EGFR-TKIs and a renewed clinical challenge of drug resistance.

[0004] Against this backdrop, this study reports a novel class of trisubstituted anisole compounds containing N-propanol indole. Experiments showed that these compounds exhibited significantly superior inhibitory activity compared to osimertinib against kinases carrying EGFR triple mutations (Del19 / T790M / C797S or L858R / T790M / C797S) and tumor cells (H1975-EGFR L858R / T790M / C797S). These results suggest that this series of compounds has the potential to be developed into fourth-generation EGFR-TKIs capable of overcoming C797S resistance mutations, providing new therapeutic hope for patients with advanced NSCLC. Summary of the Invention

[0005] The purpose of this invention is to design and synthesize a series of trisubstituted anisole compounds containing an N-propanol indole structure. In vitro activity screening showed that these compounds exhibit excellent inhibitory activity against EGFR (L858R / T790M / C797S) kinase, EGFR (Del19 / T790M / C797S) kinase, and the H1975-EGFR (L858R / T790M / C797S) cell line, and hold promise for development into novel anti-lung cancer drugs.

[0006] This invention provides trisubstituted anisole compounds with an N-propanol indole structure, having the structure shown in formula (I).

[0007] (I)

[0008] in, R1 is selected from 5-10 aryl or 5-10 heteroaryl; the heteroaryl contains 1-3 heteroatoms selected from N, O or S; the aryl or heteroaryl is substituted by 0-5 identical or different R3s; R3 is selected from hydrogen, hydroxyl, halogen, nitro, amino, cyano, morpholino, trifluoromethyl, alkyl containing 1-6 carbons, cycloalkyl containing 3-8 carbons, alkenyl containing 2-6 carbons, alkynyl containing 2-6 carbons, alkoxy containing 1-6 carbons, alkyl containing 1-6 carbons substituted with hydroxyl or amino groups or halogenated, ester group containing 1-6 carbons, alkylacyl group containing 1-6 carbons, alkylamide group containing 1-6 carbons, and hydroxymethyl group containing 1-6 carbons. R2 is selected from , , , , , , , ; X is selected from halogens and hydrogen.

[0009] Furthermore, the above-mentioned trisubstituted anisole compounds containing an N-propanol indole structure, R1 is selected from 5-10 aryl groups or 5-10 heteroaryl groups; the heteroaryl group contains 1-3 heteroatoms selected from N, O or S; the aryl group or heteroaryl group is substituted by 0-3 identical or different R3 atoms; R3 is selected from hydrogen, hydroxyl, halogen, nitro, amino, cyano, trifluoromethyl, alkyl containing 1-3 carbons, cycloalkyl containing 3-6 carbons, alkenyl containing 2-5 carbons, alkynyl containing 2-5 carbons, alkoxy containing 1-3 carbons, alkyl containing 1-3 carbons substituted with hydroxyl or amino groups or halogenated, ester group containing 1-3 carbons, alkyl acyl group containing 1-3 carbons, alkyl amide group containing 1-3 carbons, and hydroxymethyl group containing 1-3 carbons. R2 is selected from , , , , , ; X is selected from fluorine, chlorine, bromine, and hydrogen.

[0010] Furthermore, the above-mentioned trisubstituted anisole compounds containing an N-propanol indole structure, R1 is selected from 5-10 aryl groups or 5-10 heteroaryl groups; the heteroaryl group contains 1-3 heteroatoms selected from N, O or S; the aryl group or heteroaryl group is substituted by 0-3 identical or different R3 atoms; R3 is selected from hydrogen, nitro, hydroxyl, halogen, alkyl containing 1-3 carbons, alkoxy containing 1-3 carbons, ester containing 1-3 carbons, and hydroxymethyl containing 1-3 carbons; R2 is selected from , , , ; X is selected from chlorine and hydrogen.

[0011] Furthermore, the above-mentioned trisubstituted anisole compounds containing an N-propanol indole structure have the following structural formula:

[0012] A pharmaceutical composition comprising the above-described trisubstituted anisole compound containing an N-propanol indole structure and its pharmaceutically acceptable excipients.

[0013] Preferably, in the above-described pharmaceutical composition, the pharmaceutically acceptable excipient is selected from one or more of fillers, disintegrants, binders, and lubricants.

[0014] Preferably, the dosage form of the above-mentioned pharmaceutical composition is a tablet, capsule, granule, spray, or injection.

[0015] The use of any of the above-described trisubstituted anisole compounds containing an N-propanol indole structure or any of the above-described pharmaceutical compositions in the preparation of medicaments for the treatment and / or prevention of proliferative diseases.

[0016] Use in the preparation of medicaments for treating and / or preventing cancer by any of the above-described trisubstituted anisole compounds containing an N-propanol indole structure or any of the above-described pharmaceutical compositions.

[0017] The use of any of the above-described trisubstituted anisole compounds containing an N-propanol indole structure or any of the above-described pharmaceutical compositions in the preparation of drugs for the treatment and / or prevention of lung cancer.

[0018] According to some common methods in the field to which this invention pertains, the trisubstituted anisole compounds of general formula (I) of this invention containing an N-propanol indole structure can react with acids to form pharmaceutically acceptable salts, including inorganic acid addition salts and / or organic acid addition salts, wherein the acids are: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, and benzoic acid.

[0019] The beneficial effects of the present invention are as follows: (The trisubstituted anisole is mentioned in the original text.) The trisubstituted anisole compounds containing N-propanol indole structures and their pharmaceutically acceptable salts obtained in this invention have excellent inhibitory activity and antitumor activity against EGFR (L858R / T790M / C797S) kinase and EGFR (Del19 / T790M / C797S) kinase. In vitro tumor cell inhibition assays using EGFR (L858R / T790M / C797S) kinase, EGFR (Del19 / T790M / C797S) kinase, and H1975-EGFR (L858R / T790M / C797S) cell lines demonstrated that the compounds of this invention have significant inhibitory effects on EGFR (L858R / T790M / C797S) kinase, EGFR (Del19 / T790M / C797S) kinase, and corresponding tumor cells, with effects far superior to osimertinib. The compounds described in this invention are chemical pharmaceutical raw materials, specifically used for the preparation of drugs for the treatment and / or prevention of lung cancer. Detailed Implementation

[0020] The examples and preparation methods provided below further illustrate and demonstrate the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparation methods does not limit the scope of the present invention in any way.

[0021] The following synthetic route describes the preparation method of the trisubstituted anisole compound of general formula (I) of the present invention containing an N-propanol indole structure.

[0022] All raw materials were prepared by methods well known to those skilled in the art of organic chemistry, as described in the synthetic route below, or were commercially available. All final compounds of this invention were prepared by methods described in the synthetic route below or by similar methods well known to those skilled in the art of organic chemistry. All variable factors used in the synthetic route below are defined as follows or as defined above.

[0023] General synthetic route:

[0024] Reaction conditions: (i) potassium hydroxide, dimethyl sulfoxide; (ii) substituted pyrimidine, aluminum trichloride, 1,4-dioxane; (iii) 4-fluoro-2-methoxy-5-nitroaniline, 4-toluenesulfonic acid monohydrate, isopropanol; (iv) Pd / C, hydrazine hydrate, ethanol; (v) aromatic aldehyde, glacial acetic acid, 1,2-dichloroethane; (vi) amine, potassium carbonate, N,N-dimethylformamide; (vii) sodium triacetamipridoborohydride, 1,2-dichloroethane.

[0025] The synthetic routes for compounds 1-15 according to the general synthetic route of formula (I) of the present invention are as follows:

[0026] Reaction conditions: (i) potassium hydroxide, dimethyl sulfoxide; (ii) 2,4-dichloropyrimidine, aluminum trichloride, 1,4-dioxane; (iii) 4-fluoro-2-methoxy-5-nitroaniline, 4-toluenesulfonic acid monohydrate, isopropanol; (iv) Pd / C, hydrazine hydrate, ethanol; (v) aromatic aldehyde, glacial acetic acid, 1,2-dichloroethane; (vi) amine, potassium carbonate, N,N-dimethylformamide; (vii) sodium triacetamipridoborohydride, 1,2-dichloroethane.

[0027] Step A: Preparation of 3-(1H-indol-1-yl)prop-1-ol (2a)

[0028] 15 g of indole was placed in a 100 mL round-bottom flask, and 30 mL of dimethyl sulfoxide was added to dissolve it completely. 25 g of potassium hydroxide was then added to the flask. The reaction mixture was placed in an oil bath at 60 °C, and stirring was started until the solid was completely dissolved. Then, 14 g of 3-chloro-1-propanol was added dropwise using a constant-pressure titration funnel. After approximately 1.5 hours, the reaction was monitored by thin-layer chromatography to confirm completion. The reaction mixture was then removed from the oil bath and cooled. Water was added, and the mixture was extracted three times with dichloromethane. The organic phases were combined and washed once with a saturated salt solution. The mixture was then dried over anhydrous Na₂SO₄. After filtration and solvent removal, a crude solid product was obtained. Purification was performed by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give 12 g of a colorless oily intermediate 2a. MS (ESI) m / z (%): 176.2 [M+H] + .

[0029] Step B: Preparation of 3-(3-(2-chloropyrimidin-4-yl)-1H-indol-1-yl)prop-1-ol (3a)

[0030] 2,4-Dichloropyrimidine (2.0 g) and aluminum trichloride (2.2 g) were placed in a 50 mL three-necked flask, and 1,4-dioxane (5 mL) was added. The mixture was purged with nitrogen three times. The flask was placed in an oil bath at 60 °C and stirred until the starting material was completely dissolved. Intermediate 2a (2.8 g), pre-diluted with 1,4-dioxane, was added dropwise to the reaction mixture. After reacting for 3 hours, the reaction was confirmed to be complete by thin-layer chromatography. Water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined and washed with a saturated NaCl aqueous solution. The mixture was dried over anhydrous Na₂SO₄, filtered, and the solvent was evaporated to obtain a solid crude product. Purification was performed by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give 2.4 g of a white solid intermediate 3a. MS (ESI) m / z (%): 394.1 [M+H] + .

[0031] Step C: Preparation of 3-(3-(2-((4-fluoro-2-methoxy-5-nitrophenyl)amino)pyrimidin-4-yl)-1H-indol-1-yl)prop-1-ol (4a)

[0032] Intermediate 3a (2.0 g) was placed in a 50 mL three-necked flask and dissolved in 10 mL of isopropanol. Then, 4-fluoro-2-methoxy-5-nitroaniline (1.2 g) and p-toluenesulfonic acid monohydrate (4.4 g) were added. The mixture was purged with nitrogen three times, and then the flask was placed in an oil bath at 70 °C and stirred for 8 hours. After confirming the completion of the reaction by thin-layer chromatography, the reaction solution was removed from the oil bath and cooled to room temperature, where a yellow solid was observed to precipitate. To promote product precipitation, the flask was transferred to a cold trap at -20 °C and stirred for another 15 minutes. The product was then filtered using a Buchner funnel and washed thoroughly three times with isopropanol. After drying, 2.5 g of yellow solid intermediate 4a was obtained. MS (ESI) m / z (%): 438.1 [M+H] + .

[0033] Step D: Preparation of 3-(3-(2-((4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxy-5-nitrophenyl)amino)pyrimidin-4-yl)-1H-indol-1-yl)prop-1-ol (5a)

[0034] Intermediate 4a (1.3 g) and potassium carbonate solid (0.9 g) were placed in a 50 mL round-bottom flask. N,N-dimethylformamide (10 mL) was then added to the flask to completely dissolve the starting material. The flask was transferred to an oil bath at 80 °C and heated with stirring. N,N,N-trimethylethylenediamine (0.5 g) was slowly added dropwise, and the reaction was allowed to proceed for 1 hour. The reaction was monitored by thin-layer chromatography. After the reaction was complete, the reaction solution was removed from the oil bath and allowed to cool naturally to room temperature. After cooling, water was added, and the reaction solution was extracted three times with ethyl acetate. The organic phases were combined. The organic phase was then washed three times with a saturated NaCl aqueous solution. Anhydrous Na₂SO₄ was added to the organic phase for drying. After filtration, the solvent was removed by vacuum evaporation to obtain a crude solid product. Purification was performed by silica gel column chromatography (dichloromethane:methanol = 20:1) to obtain 1.2 g of red solid intermediate 5a. MS (ESI) m / z (%): 520.3 [M+H] + .

[0035] Step E: Preparation of 3-(3-(2-((2-methoxy-4-(4-methylpiperazin-1-yl)-5-nitrophenyl)amino)pyrimidin-4-yl)-1H-indol-1-yl)prop-1-ol (5b)

[0036] Using intermediate 4a (1.3 g), potassium carbonate solid (0.9 g), and 1-methylpiperazine (0.5 g) as raw materials, 1.2 g of intermediate 5b was obtained according to the method shown in step D. MS (ESI) m / z (%): 518.2 [M+H] + .

[0037] Step F: Preparation of 3-(3-(2-((4-(4-(dimethylamino)piperidin-1-yl)-2-methoxy-5-nitrophenyl)amino)pyrimidin-4-yl)-1H-indol-1-yl)prop-1-ol (5c)

[0038] Using intermediate 4a (1.3 g), potassium carbonate solid (0.9 g), and N,N-dimethylpiperidin-4-amine (0.6 g) as raw materials, 1.2 g of intermediate 5c was obtained according to the method shown in step D. MS (ESI) m / z (%): 546.3 [M+H] + .

[0039] Step G: Preparation of 3-(3-(2-((2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)-5-nitrophenyl)amino)pyrimidin-4-yl)-1H-indol-1-yl)prop-1-ol (5d)

[0040] Using intermediate 4a (1.3 g), potassium carbonate solid (0.9 g), and 1-methyl-4-(piperidin-4-yl)piperazine (0.9 g) as starting materials, 1.2 g of intermediate 5d was obtained according to the method shown in step D. MS (ESI) m / z (%): 601.3 [M+H] + .

[0041] Step H: Preparation of 3-(3-(2-((5-amino-4-((2-(dimethylamino)ethyl)(methyl)amino)-2-methoxyphenyl)amino)pyrimidin-4-yl)-1H-indol-1-yl)prop-1-ol (6a)

[0042] Intermediate 5a (2.0 g) was placed in a 50 mL round-bottom flask, and anhydrous ethanol (10 mL) and palladium on carbon (72 mg) were added. The flask was then placed in an oil bath at 70 °C. After the temperature reached 50 °C, hydrazine hydrate (5 mL) was added dropwise, and the mixture was heated to reflux at 70 °C for 45 minutes. The reaction was confirmed to be complete by thin-layer chromatography. The flask was then removed from the oil bath and cooled to room temperature. Diatomaceous earth was added and the mixture was filtered. During filtration, the filter cake was washed three times with anhydrous ethanol. The anhydrous ethanol was then removed by rotary evaporation under reduced pressure. Water was added, and the product was extracted three times with ethyl acetate. The organic phases were combined. The combined organic phases were then washed with a saturated NaCl aqueous solution. Anhydrous Na2SO4 was added to the organic phase for drying. After filtration, the solvent was removed by evaporation under reduced pressure to obtain a solid crude product. Purification was performed by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain 1.4 g of solid intermediate 6a. MS (ESI) m / z (%): 490.3 [M+H] + .

[0043] Step I: Preparation of 3-(3-(2-((5-amino-2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-1H-indol-1-yl)prop-1-ol (6b)

[0044] Using intermediate 5b (2.0 g) as a starting material, 1.3 g of solid intermediate 6b was obtained according to the method shown in step H. MS (ESI) m / z (%): 488.3 [M+H] + .

[0045] Step J: Preparation of 3-(3-(2-((5-amino-4-(4-(dimethylamino)piperidin-1-yl)-2-methoxyphenyl)amino)pyrimidin-4-yl)-1H-indol-1-yl)prop-1-ol (6c)

[0046] Using intermediate 5c (2.0 g) as a starting material, 1.5 g of solid intermediate 6c was obtained according to the method shown in step H. MS (ESI) m / z (%): 516.3 [M+H] + .

[0047] Step K: Preparation of 3-(3-(2-((5-amino-2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)-1H-indol-1-yl)prop-1-ol (6d)

[0048] Using intermediate 5d (2.0 g) as raw material, 1.6 g of solid intermediate 6d was obtained according to the method shown in step H. MS (ESI) m / z (%): 571.3 [M+H] + .

[0049] General methods for preparing compounds 1-15: 200 mg (1 equivalent) of intermediate 6a, 6b, 6c, or 6d was placed in a round-bottom flask, and 1.2 equivalents of aromatic formaldehyde were added. The solid was dissolved in 1,2-dichloroethane (10 mL), and the mixture was heated and stirred in an oil bath at 30°C. Then, 5 drops of glacial acetic acid were added dropwise. After thin-layer chromatography (TLC) detected the formation of an imine in the system, 2.5 equivalents of sodium triacetoxyborohydride were weighed and added to the reaction solution, and the reaction was continued for one hour. The reaction was monitored by TLC until completion. The pH was adjusted to 8–9 with saturated sodium carbonate solution. The reaction system was then transferred to a separatory funnel, and the aqueous phase was extracted three times with dichloromethane. The organic phase was collected and washed with saturated sodium chloride solution. Anhydrous sodium sulfate was added to the organic phase to dehydrate and dry it. After filtration, the mother liquor was distilled under reduced pressure to obtain the solid product. The crude product was separated and purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to finally obtain solid product compounds 1–15.

[0050] Compounds 1 to 15 were prepared using the general preparation method described above (see Table 1).

[0051] Table 1. Structural formulas and 1H NMR and mass spectrometry data of compounds 1-5

[0052]

[0053]

[0054] Example 16: Assay for the EGFR kinase inhibitory activity of the compound

[0055] Operating steps: (1) Prepare a kinase buffer of 1 volume: Mix 50 mM 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES) (pH = 7.5), 10 mM MgCl2, 4 mM dithiothreitol (DTT), 0.01% Tween-20 (v / v) and 0.01% bovine serum albumin (BSA).

[0056] (2) Preparation of compounds: serial dilution of compounds and preparation of blank controls

[0057] a. Dissolve compounds 1–15 and osimertinib in 100% dimethyl sulfoxide (DMSO) and dilute to 100-fold the final desired maximum inhibitory concentration. For example, if the compounds are tested at 1 μM, then prepare a 100 μM DMSO solution of the compounds.

[0058] b. Add 100 μL of 100% DMSO to two empty wells in the same 96-well plate without a composite control or enzyme control. Label this plate as a blank control.

[0059] c. Transfer 40 μL of the compound from the blank control to a new 384-well plate as an intermediate plate.

[0060] d. Prepare the test plate: Repeatedly transfer 200 nL per well from the 384-well plate to the 384-well test plate. For example, A1 in the 384-well plate is transferred to A1 and A2 in another 384-well plate, A2 in the 384-well plate is transferred to A3 and A4 in another 384-well plate, and so on.

[0061] (3) Kinase response

[0062] a. Prepare 2x kinase solution: Prepare kinase solution in 1x kinase buffer at a concentration twice the final concentration of each reagent; add 10 μL of kinase solution (or 5 μL of 1x kinase buffer) to each well of the test plate, except for the control wells which do not contain enzyme; shake the plate and incubate at room temperature for a period of time.

[0063] b. Prepare 2x substrate solutions: Prepare substrate solutions of luciferin-polyGT and ATP in 1x kinase reaction buffer at a concentration twice the final reagent concentration required for each experiment; add 10 μL of substrate solution to each well of the assay plate and start the reaction; shake the plate.

[0064] c. Cover the experimental plate and incubate at room temperature for the specified time.

[0065] (4) Kinase detection

[0066] a. Prepare a detection solution with a final concentration of 2x in antibody dilution buffer; b. Add 20 μL of detection solution to each well of the detection plate and stop the reaction; c. Briefly mix in a centrifuge, incubate at room temperature for the specified time, and then perform fluorescence reading on a plate reader.

[0067] (5) Data reading: Data is collected on the Envision program with an excitation wavelength of 340 nm and an emission wavelength of 520 nm and 495 nm.

[0068] (6) Calculate the inhibition rate: Copy the RFU value from the Envision program; calculate the ratio of RFU 520 nm / RFU 495 nm; convert the ratio to an inhibition percentage value, inhibition percentage = (maximum sample ratio) / (maximum and minimum values) 100, where "min" represents the ratio controlled without enzyme and "max" represents the ratio controlled by DMSO. Data are presented in MS Excel, and curves were fitted using XLFit Excel. IC 50 The formula for calculating the half-maximal inhibitory concentration (IC50) is: Y = Bottom + (Top - Bottom) / (1 + (IC50) / 2) 50 / X)^HillSlope

[0069] The activity data of the compounds in inhibiting EGFR (L858R / T790M / C797S) kinase and EGFR (Del19 / T790M / C797S) kinase are shown in Table 2: IC in Table 2 50 ≤ 10 nM, denoted by A; 10 nM < IC 50 ≤ 50 nM, denoted by B; 50 nM < IC 50 ≤ 100 nM, denoted by C; 100 nM < IC 50 ≤ 150 nM, denoted by D; IC 50 >150 nM, denoted by E.

[0070] Table 2. Activity data of compounds 1-15 and osimertinib inhibiting EGFR (L858R / T790M / C797S) and EGFR (Del19 / T790M / C797S) kinases.

[0071] As shown in Table 2, the trisubstituted anisole compounds 1-15 containing the N-propanol indole structure exhibit excellent inhibitory activity against both EGFR (L858R / T790M / C797S) and EGFR (Del19 / T790M / C797S) kinases. In particular, their inhibitory activity against EGFR (L858R / T790M / C797S) kinase is more than 15 times that of the control drug osimertinib. This class of compounds shows promising potential for the development and application of antitumor drugs.

[0072] Example 17: In vitro antitumor cell proliferation activity test of the compound

[0073] The in vitro inhibitory activity of the trisubstituted anisole compounds containing the N-propanol indole structure of this invention on the H1975-EGFR (L858R / T790M / C797S) cell line was tested using the CTG method.

[0074] Operating steps: Dissolve the test compounds 1-15 and osimertinib separately in DMSO to a stock solution concentration of 20 mM. Resuspend the cells in culture medium and adjust their density to the recommended value. Add 90 μL of cell suspension to the assay plate, followed by culture medium. Incubate the cells overnight at 37°C, 5% CO2, and 100% relative humidity. Prepare a composite stock solution plate (200X stock solution plate) by sequentially diluting the stock solution in DMSO from the highest to the lowest concentration. Dilute the 200X stock solution to 10X with culture medium, add 10 μL of the compound to each well, and normalize the final DMSO concentration to 0.5%. Incubate for 3 days. Equilibrate the assay plate and CellTiter Glo reagent at room temperature for approximately 30 minutes, then add 50 μL of CellTiter Glo reagent to each well. Shake for 2 minutes, incubate at room temperature for 10 minutes to stabilize the luminescence signal, and then detect using EnVision. The inhibition rate (IR) of the measured compound is: IR(%) = (1 - (RLU compound - RLU blank) / (RLU control - RLU blank)) 100%. The data was interpreted and the corresponding IC was calculated using GraphPadPrism. 50 (Half-maximal inhibitory concentration) value. The inhibition results on the H1975-EGFR (L858R / T790M / C797S) cell line are shown in Table 3.

[0075] IC in Table 3 50 ≤ 3.5 μM, denoted by A; 3.5 μM < IC 50 ≤ 4.5 μM, denoted by B; IC 50 >4.5 μM, denoted by C.

[0076] Table 3. Inhibition results of compounds 1-15 and osimertinib on the H1975-EGFR (L858R / T790M / C797S) cell line.

[0077] As shown in Table 3, the trisubstituted anisole compounds containing the N-propanol indole structure exhibit excellent inhibitory activity against the H1975-EGFR (L858R / T790M / C797S) cell line, significantly higher than that of osimertinib, a third-generation EGFR inhibitor. These data indicate that trisubstituted anisole compounds containing the N-propanol indole structure have promising prospects for development and application as antitumor drugs.

[0078] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details of the above embodiments. Within the scope of the technical concept of the present invention, various equivalent transformations can be made to the technical solutions of the present invention. To avoid unnecessary repetition, the present invention will not describe the various possible combinations separately. Any modifications, equivalent substitutions, or improvements made within the scope of the technical concept of the present invention are included within the protection scope of the present invention.

Claims

1. Trisubstituted anisoles containing an N-propylol-indole structure, characterized in that, It has the following structural formula: 。 2. A pharmaceutical composition, characterized by comprising: This includes the trisubstituted anisole compounds containing the N-propanol indole structure as described in claim 1, and pharmaceutically acceptable excipients thereof.

3. The pharmaceutical composition of claim 2, wherein The pharmaceutically acceptable excipient is selected from one or more of fillers, disintegrants, binders, and lubricants.

4. The pharmaceutical composition of claim 2, wherein The dosage form of the pharmaceutical composition is tablets, capsules, granules, sprays, or injections.

5. The use of the trisubstituted anisole compound containing an N-propanol indole structure according to claim 1 or the pharmaceutical composition according to any one of claims 2-4 in the preparation of a medicament for treating and / or preventing proliferative diseases.

6. The use of the trisubstituted anisole compound containing an N-propanol indole structure according to claim 1 or the pharmaceutical composition according to any one of claims 2-4 in the preparation of a medicament for treating and / or preventing cancer.

7. The application according to claim 6, characterized in that, The cancer mentioned is lung cancer.

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