Synthesis of 4-adamantyl-based 3-methylphenol derivative and antitumor activity research of 4-adamantyl-based 3-methylphenol derivative

Abstract

The invention discloses synthesis and antitumor activity research of a novel drug small molecular formula (1) containing an adamantane structure. R is 2-[4-(1-adamantyl)-3-methylphenoxy] acetic acid obtained by taking 1-bromoadamantane as a starting raw material and then carrying out free radical substitution, oxidation reaction and demethylation reaction; secondly, carrying out condensation reaction on the 2-[4-(1-adamantyl)-3-methylphenoxy] acetic acid and amino to obtain a target compound. A target compound is subjected to a CCK-8 experiment, and a conclusion is obtained that the target compound 7a-7h and 5-fluorouracil are tested at the concentration of 20 [mu] M, and it can be known that 7g of the compound has a good HeLa cell inhibition effect, and 7c of the compound has the best inhibition effect on A549 cells and Hep G2 cells. IC50 value calculation is carried out on a target compound under various concentrations, the IC50 value of 7g on HeLa cells is minimum, the inhibiting effect is best, and the IC50 value of 7c on A549 cells and Hep G2 cells is minimum, and the inhibiting effect is best.

Description

TECHNICAL FIELD

[0001] The present application relates to the synthesis of a small molecule drug containing adamantane structure and the research on its anti-tumor activity. BACKGROUND

[0002] Cancer is considered the second largest disease that is fatal. According to the Global Cancer Report 2021 published by the World Health Organization (WHO) on February 3, 2021, the global cancer incidence and mortality are growing at an alarming rate, and on average, 1 out of 8 deaths is caused by cancer. The report predicts that the global cancer cases will continue to grow from 14 million people in 2021 to 19 million people in 2025, and to 24 million people in 2035. In developing and developed countries, lung cancer, breast cancer, colon cancer and melanoma are the most common types of cancer. And in developed countries, lung cancer and breast cancer are more common. Among all the current treatment methods, chemotherapy and drug assistance are still the most common choices for cancer treatment, but most anticancer drugs are hindered by serious toxic side effects. Therefore, the development of new therapeutic drugs will have important research significance.

[0003] Adamantane is mainly used for the synthesis of anticancer, antitumor and other special drugs, which is an alicyclic hydrocarbon with good liposolubility, and the skeleton of adamantane is non-toxic, which can change its pharmacological activity. According to previous studies, the introduction of amino group into the molecule can improve the water solubility and enhance the anticancer activity. Because different substituents have different physicochemical properties, the properties of the target compounds are determined, and the inhibition of tumor cells will also have different effects. The research of these compounds will provide valuable information for the synthesis and research of new small molecule anticancer inhibitors. SUMMARY

[0004] The present application discloses a preparation method of a small molecule drug containing adamantane structure, and uses CCK-8 method to conduct toxicity experiment on cells to evaluate the anti-proliferation effect of the compound on A549, HeLa, Hep G2 and normal mouse fibroblast L929.

[0005] The present application adopts the following technical scheme, a small molecule drug containing adamantane structure, the structural formula is:

[0006] In the present application, R is R1, R2 and R3 are each independently H, OCH3 or COOCH3.

[0007] The present application synthesizes 2-[4-(1-adamantyl)-3-methylphenylamino] acetic acid through typical Friedel-Crafts alkylation reaction, Wittig reaction and hydrolysis reaction, etc.

[0008] In the present application, 2-[4-(1-adamantyl)-3-methylphenylamino] acetic acid and different amine compounds are coupled to obtain a drug small molecule containing adamantane structure: formula (1).

[0009] The present application is characterized by nuclear magnetic resonance, high resolution mass spectrometry, infrared spectroscopy, etc., and the results fully confirm the synthesis of the compound. BRIEF DESCRIPTION OF DRAWINGS

[0010] Figure 1 is a synthesis route; Figure 2 is a target product of the synthesis route; Figure 3 is a hydrogen spectrum of an intermediate compound; Figures 4-11 is a hydrogen spectrum and carbon spectrum of the target compound; Figure 12 is the inhibition rate of the target compound and 5-fluorouracil on different cancer cells at a concentration of 20 micromoles; Figure 13 is an IC 50 value graph of the target compound; DETAILED DESCRIPTION

[0011] The present application will be specifically and in detail described below. The present embodiment is implemented on the premise of the technical scheme of the present application, and detailed implementation modes and specific operation processes are given. It should be noted that, for ordinary skilled persons in the technical field, several improvements and refinements can be made without departing from the principle of the present application, and these improvements and refinements should also be regarded as the protection scope of the present application.

[0012] Example 1: Synthesis of 2-[4-(1-adamantyl)-3-methylphenoxy] acetic acid (1) Synthesis of (3R, 5R, 7R)-2-[4-(l-adamantyl)-3-methylphenoxy]ethyl acetate (intermediate compound 4) Take 100 mL single neck flask, at room temperature, add 4-[(3r, 5r, 7r)-adamant-l-yl]-3-methylphenol 0.1 g, add cesium carbonate 0.3 g, add a certain amount of N, N-dimethylformamide to dissolve, then add 100 μL of methyl chloroacetate, transfer the whole reaction system to a 60 degree Celsius oil bath, connect a straight condenser tube, introduce nitrogen, time, and react for 6 hours. After the reaction is completed, cool to room temperature, concentrate, and elute by column chromatography using an eluent (V petroleum ether: V dichloromethane = 1:1). After elution, spin off the solvent using a rotary evaporator, and dry the residue in a vacuum drying oven to obtain a white solid. Yield: 55%. (2) Synthesis of 2-[4-(l-adamantyl)-3-methylphenoxy]acetic acid Take 100 mL single neck flask, at room temperature, add (3R, 5R, 7R)-2-[4-(l-adamantyl)-3-methylphenoxy]ethyl acetate 0.1 g, add sodium hydroxide 1.5 g, add water 10 mL, and a certain amount of anhydrous ethanol as a reaction solvent, transfer the whole reaction system to an 80 degree Celsius oil bath, connect a condenser tube, protect with nitrogen, and reflux for 4 hours. After the reaction is completed, cool to room temperature, remove the ethanol after removing the rotor, add diluted concentrated hydrochloric acid, and make the pH value acidic to precipitate a flocculent precipitate. Filter, dry the filter cake in a vacuum drying oven overnight to obtain a white solid. Yield: 88%. 1H NMR (400 MHz, CDC13) δ 7.14 (d, J = 7.9 Hz, 1H), 6.81 (d, J = 7.8 Hz, 1H), 6.57 (s, 1H), 4.69 (s, 2H), 2.31 (s, 3H), 2.10 (s, 6H), 2.07 (s, 3H), 1.77 (s, 6H).

[0013] Example 2: Synthesis of target compounds 7a-7h (1) (3R, 5R, 7R)-2-[4-(l-adamantyl)-3-methylphenoxy]-N-phenylacetamide (compound 7a) Take 100 mL single mouth bottle, at room temperature, add 2-[4-(1- adamantyl)-3-methylphenoxy] acetic acid 0.1 g, add 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide (EDC) 0.1 g, 1-hydroxybenzotriazole (HOBt) 0.1 g, N, N- Diisopropylethylamine (DIPEA) 116 μL, add a certain amount of dichloromethane and super dry N, N-dimethylformamide to dissolve it, transfer the whole reaction system to a magnetic stirrer, protect it with nitrogen, and react at room temperature for 1 hour. After 1 hour, add aniline 60 μL, continue to react for 12 hours. After the reaction is completed, wash with saturated NaCl solution and dichloromethane for several times, dry with anhydrous magnesium sulfate for one hour, after drying, filter, concentrate the filtrate, use eluent (V dichloromethane: V ethyl acetate = 20: 1) to elute by column chromatography, remove the solvent after elution, place the residue in a vacuum drying oven to dry overnight, and obtain white solid. Yield: 43%. 1H NMR (400 MHz, CDC13) δ 7.56 (d, J = 7.5 Hz, 2H), 7.37 (s, 2H), 7.19 (d, J = 8.0 Hz, 2H), 6.85 (d, J = 8.2 Hz, 1H), 6.71 (s, 1H), 4.67 (s, 2H), 2.31 (s, 3H), 2.15 (s, 9H), 1.79 (s, 6H).

[0014] (2) Synthesis of (3R, 5R, 7R)-2-[4-(1-adamantyl)-3-methylphenoxy]-N-(2- methoxyphenyl)acetamide (compound 7b) Take 100 mL single mouth bottle, at room temperature, add 2-[4-(1-adamantyl)-3- methylphenoxy] acetic acid 0.1 g, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) 0.1 g, 1-hydroxybenzotriazole (HOBT) 0.1 g, N, N-Diisopropylethylamine (DIPEA) 116 μL, add a certain amount of dichloromethane and super dry N, N-dimethylformamide to dissolve it, transfer the whole reaction system to a magnetic stirrer, protect it with nitrogen, and react at room temperature for 1 hour. After 1 hour, add aniline 60 μL, continue to react for 12 hours. After the reaction is completed, wash with saturated NaCl solution and dichloromethane for several times, dry with anhydrous magnesium sulfate for one hour, after drying, filter, concentrate the filtrate, use eluent (V dichloromethane: V ethyl acetate = 20: 1) to elute by column chromatography, remove the solvent after elution, place the residue in a vacuum drying oven and dry overnight to obtain white solid. Yield: 43%. 1H NMR (400 MHz, CDCl3) δ 8.93 (s, 1H), 8.43 (d, J = 7.9 Hz, 1H), 7.18 (d, J = 7.9 Hz, 1H), 7.09 (t, J = 7.8 Hz, 1H), 7.00 (t, J = 7.7 Hz, 1H), 6.86 (dd, J = 15.0, 8.0 Hz, 2H), 6.73 (s, 1H), 4.69 (s, 2H), 3.78 (s, 3H), 2.30 (s, 3H), 2.15 (s, 6H), 2.09 (s, 3H), 1.77 (s, 6H).

[0015] (3) Synthesis of (3R, 5R, 7R)-2-[4-(1-adamantyl)-3-methylphenoxy]-N-(3- methoxyphenyl)acetamide (compound 7c) Take 100 mL single mouth bottle, at room temperature, add 2-[4-(1-adamantyl)-3- methylphenoxy] acetic acid 0.1 g, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) 0.1 g, 1-hydroxybenzotriazole (HOBT) 0.1 g, N, N-Diisopropylethylamine (DIPEA) 116 μL, add a certain amount of dichloromethane and super dry N, N-dimethylformamide to dissolve it, transfer the whole reaction system to a magnetic stirrer, protect it with nitrogen, and react at room temperature for 1 hour. After 1 hour, add aniline 60 μL and continue to react for 12 hours. After the reaction is completed, wash with saturated NaCl solution and dichloromethane several times, dry with anhydrous magnesium sulfate for one hour, after drying, filter, concentrate the filtrate, use eluent (V dichloromethane: V ethyl acetate = 20: 1) to elute by column chromatography, remove the solvent after elution, and dry the residue in a vacuum drying oven overnight to obtain a white solid. Yield: 48%. 1H NMR (400 MHz, CDCl3) δ 8.31 (s, 1H), 7.30 (d, J = 2.4 Hz, 1H), 7.27 (d, J = 5.4 Hz, 1H), 7.19 (dd, J = 8.0, 2.3 Hz, 1H), 7.05 (d, J = 7.2 Hz, 1H), 6.86 (d, J = 8.0 Hz, 1H), 6.71 (s, 1H), 4.66 (d, J = 2.4 Hz, 2H), 3.82 (d, J = 2.4 Hz, 3H), 2.32 (d, J = 2.4 Hz, 3H), 2.15 (s, 9H), 1.80 (s, 6H).

[0016] (4) (3R, 5R, 7R)-2-[4-(1-adamantyl)-3-methylphenoxy]-N-(4-methoxyphenyl)acetamide (compound 7d) Take 100 mL single mouth bottle, at room temperature, add 2-[4-(1-adamantyl)-3- methylphenoxy] acetic acid 0.1 g, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) 0.1 g, 1-hydroxybenzotriazole (HOBT) 0.1 g, N, N-Diisopropylethylamine (DIPEA) 116 μL, add a quantitative amount of dichloromethane and super dry N, N-dimethylformamide to dissolve it, transfer the whole reaction system to a magnetic stirrer, protect it with nitrogen, and react at room temperature for 1 hour. After 1 hour, add aniline 60 μL, continue to react for 12 hours. After the reaction is completed, wash with saturated NaCl solution and dichloromethane several times, dry with anhydrous magnesium sulfate for one hour, after drying, filter, concentrate the filtrate, use eluent (V dichloromethane: V ethyl acetate = 20: 1) to elute by column chromatography, remove the solvent after elution, and place the residue in a vacuum drying oven to dry overnight to obtain a white solid. Yield: 39%.1H NMR (400 MHz, CDCl3) δ 7.46 (d, J = 9.0 Hz, 2H), 7.18 (d, J = 7.9 Hz, 1H), 6.90 (d, J = 9.0 Hz, 2H), 6.84 (d, J = 7.8 Hz, 1H), 6.71 (s, 1H), 4.66 (s, 2H), 3.81 (s, 3H), 2.31 (s, 3H), 2.14 (s, 6H), 2.12 (s, 3H), 1.79 (s, 6H).

[0017] (5) (3R, 5R, 7R)-3-{[2-(4-(1-adamantyl)-3-methylphenoxy)acetyl]amino}benzoic acid methyl ester (compound 7e) Take 100 mL single mouth bottle, at room temperature, add 2-[4-(1-adamantyl)-3- methylphenoxy] acetic acid 0.1 g, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) 0.1 g, 1-hydroxybenzotriazole (HOBT) 0.1 g, N, N-Diisopropylethylamine (DIPEA) 116 μL, add a certain amount of dichloromethane and super dry N, N-dimethylformamide to dissolve it, transfer the whole reaction system to a magnetic stirrer, protect it with nitrogen, and react at room temperature for 1 hour. After 1 hour, add aniline 60 μL, continue to react for 12 hours. After the reaction is completed, wash with saturated NaCl solution and dichloromethane several times, dry with anhydrous magnesium sulfate for one hour, after drying, filter, concentrate the filtrate, use eluent (V dichloromethane: V ethyl acetate = 20:1) to elute by column chromatography, remove the solvent after elution, place the residue in a vacuum drying oven and dry overnight to obtain a white solid. Yield: 48%.1H NMR (400 MHz, CDCl3) δ 8.07 (s, 1H), 7.96 (d, J = 7.9 Hz, 1H), 7.84 (d, J = 7.9 Hz, 1H), 7.47 (d, J = 7.9 Hz, 1H), 7.23-7.17 (m, 1H), 6.87 (s, 1H), 6.71 (s, 1H), 4.68 (s, 2H), 3.92 (s, 3H), 2.32 (s, 3H), 2.17 (d, J = 13.7 Hz, 9H), 1.80 (s, 6H).

[0018] (6) (3R, 5R, 7R)-4-{[2-(4-(1-adamantyl)-3-methylphenoxy)acetyl]amino}benzoic acid methyl ester (compound 7f) Take 100 mL single mouth bottle, at room temperature, add 2-[4-(1-adamantyl)-3- methylphenoxy] acetic acid 0.1 g, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) 0.1 g, 1-hydroxybenzotriazole (HOBT) 0.1 g, N, N-Diisopropylethylamine (DIPEA) 116 μL, add a quantitative amount of dichloromethane and super dry N, N-dimethylformamide to dissolve it, transfer the whole reaction system to a magnetic stirrer, protect it with nitrogen, and react at room temperature for 1 hour. After 1 hour, add aniline 60 μL and continue to react for 12 hours. After the reaction is completed, wash with saturated NaCl solution and dichloromethane several times, dry with anhydrous magnesium sulfate for one hour, after drying, filter, concentrate the filtrate, use eluent (V dichloromethane: V ethyl acetate = 20:1) to elute by column chromatography, remove the solvent after elution, place the residue in a vacuum drying oven and dry overnight to obtain a white solid. Yield: 45%.1H NMR (400 MHz, CDCl3) δ 8.05 (d, J = 8.7 Hz, 2H), 7.65 (d, J = 8.8 Hz, 2H), 7.19 (d, J = 7.9 Hz, 1H), 6.86 (d, J = 8.0 Hz, 1H), 6.70 (s, 1H), 4.68 (s, 2H), 3.91 (s, 3H), 2.31 (s, 3H), 2.14 (s, 9H), 1.78 (s, 3H).

[0019] (7) (3R, 5R, 7R)-2-[4-(1-adamantyl)-3-methylphenoxy]-N-(3-cyanophenyl)acetamide (Compound 7f) Take 100 mL single mouth bottle, at room temperature, add 2-[4-(1-adamantyl)-3- methylphenoxy] acetic acid 0.1 g, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) 0.1 g, 1-hydroxybenzotriazole (HOBT) 0.1 g, N, N-Diisopropylethylamine (DIPEA) 116 μL, add a certain amount of dichloromethane and super dry N, N-dimethylformamide to dissolve it, transfer the whole reaction system to a magnetic stirrer, protect it with nitrogen, and react at room temperature for 1 hour. After 1 hour, add aniline 60 μL, continue to react for 12 hours. After the reaction is completed, wash with saturated NaCl solution and dichloromethane for several times, dry with anhydrous magnesium sulfate for one hour, after drying, filter, concentrate the filtrate, use eluent (V dichloromethane: V ethyl acetate = 20:1) to elute by column chromatography, remove the solvent after elution, place the residue in a vacuum drying oven to dry overnight, and obtain white solid. Yield: 47%.1H NMR (400 MHz, CDCl3) δ 8.41 (s, 1H), 8.03 (s, 1H), 7.69 (s, 1H), 7.46 (s, 2H), 7.19 (d, J = 8.0 Hz, 1H), 6.87 (d, J = 9.8 Hz, 1H), 6.69 (s, 1H), 4.69 (s, 2H), 2.31 (s, 3H), 2.13 (s, 9H), 1.82-1.73 (m, 6H).

[0020] (8) (3R, 5R, 7R)-2-[4-(1-adamantyl)-3-methylphenoxy]-N-[(furan-2-yl)methyl]acetamide (Compound 7f) Take 100 mL single mouth bottle, at room temperature, add 2-[4-(1-adamantyl)-3-methyl phenoxy] acetic acid 01 g, add 1-(3-dimethyl aminopropyl)-3-ethyl carbodiimide (EDC) 01 g, 1-hydroxybenzotriazole (HOBT) 01 g, N, N-diisopropyl ethylamine (DIPEA) 116 μL, add a quantitative amount of dichloromethane and super dry N, N-dimethylformamide to dissolve it, transfer the whole reaction system to a magnetic stirrer, protect it with nitrogen, and react at room temperature for 1 hour. After 1 hour, add aniline 60 μL and continue to react for 12 hours. After the reaction is completed, wash with saturated NaCl solution and dichloromethane several times, dry with anhydrous magnesium sulfate for one hour, after drying, filter, concentrate the filtrate, use the eluent (V dichloromethane: V ethyl acetate = 20:1) to elute by column chromatography, remove the solvent after elution, and dry the residue in a vacuum drying oven overnight to obtain a white solid. Yield: 49%. 1H NMR (400 MHz, CDCl3) δ 7.34 (s, 1H), 7.13 (d J = 8.0 Hz, 1H), 6.96 (s, 1H), 6.81 (d J = 8.0 Hz, 1H), 6.63 (s, 1H), 6.32 (s, 1H), 6.25 (s, 1H), 4.61-4.54 (m, 4H), 2.29 (s, 3H), 2.05 (s, 9H), 1.78-1.65 (m, 6H).

[0021] Example 3: Results of in vitro cytotoxicity evaluation The present application carries out CCK-8 experiment test on target compounds 7a-8h, expresses HeLa, A549, Hep G2 and L929, and tests target compounds 7a-8h and 5-fluorouracil at a concentration of 20 μM. As can be seen from the figure, 7c containing the structure of the amine compound with methoxy grafted at the end of the benzene ring of the adamantane structure has the best inhibition effect on A549 cells and Hep G2 cells, with an inhibition rate of 59.73% and 49.73%, while 5-fluorouracil has an inhibition rate of 39.64% on A549 cells and 40.85% on Hep G2 cells. 7g containing the structure of the amine compound with nitrile group grafted at the end of the benzene ring of the adamantane structure has the best inhibition effect on HeLa cells, with an inhibition rate of 61.77%, while 5-fluorouracil has an inhibition rate of 39.48% on HeLa cells

[0022] The present application calculates the IC 50 values of compounds 7a-7h at concentrations of 40 μM, 20 μM, 10 μM and 5 μM on HeLa, A549, Hep G2 and L929 cells, wherein 7g has a lower IC 50 value on HeLa cells, and 7c has a lower IC 50The values are low. For the amine compounds in which the benzene ring ends are connected to different groups at the 3rd position, the compounds containing methoxy groups are significantly more effective than the compounds containing methyl benzoate in A549 cells and HepG2 cells. However, in HeLa cells, the compounds containing nitrile groups are more effective than the compounds containing methyl benzoate, and all of the target compounds are non-toxic to L929 cells. Figure 11 IC50 values of target compounds 7a-7h against different cells 50 values.

Claims

1. A novel adamantane-containing drug-like small molecule (1) characterized by, ###00001### 1 The molecular chemical structure is as follows, 2. R is R1, R2, R3 are H, OCH3, or COOCH3 in the corresponding position.

3. The adamantane derivative of formula (1) as claimed in claim 1, characterized in that, Target product:

4. The adamantane derivative of formula (1) as claimed in claim 1, characterized in that, Synthetic route:

5. The use of adamantane structural derivative formula (1) having anti-tumor and other biological activities.

Images