Synthesis of adamantane drug micromolecules modified by different groups and antitumor property research of adamantane drug micromolecules modified by different groups

Abstract

The invention discloses synthesis of an adamantane structure drug small molecular formula (1) modified by different groups and anti-tumor property research # imgabs0 #, wherein R is # imgabs1 #. 1-bromoadamantane is used as an initial raw material, and 5-(adamantane-1-yl)-2-hydroxyphenol formaldehyde [1] is generated through typical Friedel-Crafts alkylation reaction, NBS bromination reaction and the like; synthesizing an important intermediate compound 5 through Wittig reaction, and carrying out coupling reaction on the intermediate compound 5 and different acyl chloride amine compounds to obtain five target compounds 7a-7e: formula (1). A CCK-8 experiment is carried out on a target compound to obtain a conclusion that the target compound 7a-7f and 5-fluorouracil are tested at the concentration of 20 mu M, and the result shows that the compound 7c has the best inhibition effect on HepG2 and HeLa cells and the compound 7e has the better inhibition effect on A549 cells. IC50 value calculation is carried out on a target compound under various concentrations, the IC50 value of 7c on HepG2 and HeLa cells is minimum and the inhibition effect is the best, and the IC50 value of 7e on A549 cells is minimum and the inhibition effect is the best.

Description

Technical Field

[0001] The present invention relates to the synthesis of a small molecule of adamantane drugs modified with different groups and the study of their anti-tumor properties. Background Art

[0002] Cancer refers to all malignant tumors. Lung cancer has become the first major cancer among new cancers, followed by colorectal cancer, gastric cancer, and breast cancer. Cancer is one of the major diseases that seriously threaten human life. Among all the current treatment methods, traditional tumor treatment methods include surgical treatment, radiotherapy, chemotherapy, traditional Chinese medicine treatment, endocrine therapy, etc. Clinically, a variety of comprehensive treatment methods are generally used. Drug therapy, as one of the main treatments for tumors, plays a major role in cancer treatment, especially for patients with tumor metastasis. However, most anticancer drugs are hindered by serious toxic side effects. Therefore, the development of new therapeutic drugs will be of great research significance.

[0003] Adamantane is a class of compounds with special spatial structures and good lipophilicity. Its derivatives have shown a variety of pharmacological activities in clinic, such as antiviral, immune enhancement and anti-tumor. In recent years, the introduction of adamantane as a skeleton into anti-tumor compounds can effectively improve the affinity of the compounds with the active pockets around the recognition area on the receptor surface, and enhance their activity and selectivity against tumor cells. The study of these compounds will provide valuable information for the synthesis and research of new small molecule anticancer inhibitors. Summary of the invention

[0004] 1. The present invention discloses a method for preparing a small drug molecule containing an adamantane structure, and uses the CCK8 method to conduct a toxicity experiment on cells to evaluate the anti-proliferation effect of the compound on A549, HeLa, HepG2 and normal mouse fibroblast L929.

[0005] 2. The present invention adopts the following technical scheme, containing a small molecule of adamantane structure drug, the structural formula is

[0006] 3. In the present invention, R is R1, R2, and R3 are H, OCH3, and CH3, respectively.

[0007] 4. The present invention uses 1-bromoadamantane as a starting material, and synthesizes 5-(adamantan-1-yl)-2-hydroxyphenol formaldehyde through typical Friedel-Crafts alkylation reaction, NBS bromination reaction, Wittig reaction and other multi-step reactions, and the structural formula is:

[0008] 5. In the present invention, 5-(adamantan-1-yl)-2-hydroxyphenol formaldehyde and different amine compounds are coupled to obtain a small drug molecule containing an adamantane structure: formula (1).

[0009] 6. The present invention 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 THE DRAWINGS

[0010] Figure 1 It is a synthetic route.

[0011] Figure 2 It is the target product of the synthetic route.

[0012] Figure 3-7 It is the hydrogen spectrum of intermediate compound 6a-6e.

[0013] Figure 8-17 These are the hydrogen and carbon spectra of 7a-7e.

[0014] Fig.18 It is the inhibition rate of target compounds 7a-7e and 5-fluorouracil on different cancer cells at a concentration of 20 μM.

[0015] Fig.19 IC50 values ​​of target compounds 7a-7f for different cells DETAILED DESCRIPTION

[0016] Example 1: Synthesis of Intermediates 6a-6e (1) Synthesis of 2-chloro-N-phenylacetamide (Intermediate Compound 6a) Take a 50ml single-mouth bottle, add 200μL of aniline, add 435.23μL of triethylamine, add a certain amount of dichloromethane to dissolve it, add 485.76μL of chloroacetyl chloride, and stir at room temperature for 4 hours. Extract with dichloromethane and sodium chloride solution, dry the organic phase with anhydrous magnesium sulfate, concentrate the filtrate, use eluent (V dichloromethane) to elute through column chromatography, after elution, spin out the solvent with a rotary evaporator, and place the residue in a vacuum drying oven to dry to obtain a white solid. Yield: 80%. 1 HNMR (400MHz, CDCl3) δ8.25 (s, 1H), 7.58 (d, J = 7.8Hz, 2H), 7.39 (t, J = 7.8Hz, 2H), 7.21 (t, J = 7.4Hz, 1H), 4.23 (s, 2H).

[0017] (2) Synthesis of 2-chloro-N-(3-methoxyphenyl)-acetamide (Intermediate Compound 6b) Take a 50ml single-mouth bottle, add 200μL of m-methoxyaniline, add 328.65μL of triethylamine, add a certain amount of dichloromethane to dissolve it, add 366.81μL of chloroacetyl chloride, and stir at room temperature for 4 hours. Extract with dichloromethane and sodium chloride solution, dry the organic phase with anhydrous magnesium sulfate, concentrate the filtrate, use eluent (V dichloromethane) to elute through column chromatography, after elution, spin out the solvent with a rotary evaporator, and place the residue in a vacuum drying oven to dry to obtain a white solid. Yield: 70%. 1H NMR (400MHz, CDCl3) δ 8.94 (s, 1H), 8.34 (d, J = 8.0Hz, 1H), 7.10 (t, J = 7.8Hz, 1H), 6.98 (t, J = 7.7Hz, 1H), 6.91 (d, J = 8.1Hz, 1H), 4.19 (s, 2H), 3.91 (s, 3H).

[0018] (3) Synthesis of methyl 2-(2-chloroethaneylamino)benzoate (intermediate compound 6c) Take a 50ml single-mouth bottle, add 200μL of methyl anthranilate, add 267.76μL of triethylamine, add quantitative dichloromethane to dissolve it, add 298.86μL of chloroacetyl chloride, and stir at room temperature for 4 hours. Extract with dichloromethane and sodium chloride solution, dry the organic phase with anhydrous magnesium sulfate, concentrate the filtrate, use eluent (V dichloromethane) to elute through column chromatography, after elution, spin out the solvent with a rotary evaporator, and place the residue in a vacuum drying oven to dry to obtain a white solid. Yield: 75.3%. 1H NMR (400MHz, CDCl3) δ11.90 (s, 1H), 8.71 (d, J = 8.5Hz, 1H), 8.07 (d, J = 7.9Hz, 1H), 7.57 (d, J = 8.4Hz, 1H), 7.17 (d, J = 7.7Hz, 1H), 4.22 (s, 2H), 3.96 (s, 3H).

[0019] (4) Synthesis of methyl 3-(2-chloroethaneylamino)benzoate (intermediate compound 6d) Take a 50ml single-mouth bottle, add 200μL of methyl m-aminobenzoate, add 267.76μL of triethylamine, add quantitative dichloromethane to dissolve it, add 298.86μL of chloroacetyl chloride, and stir at room temperature for 4 hours. Extract with dichloromethane and sodium chloride solution, dry the organic phase with anhydrous magnesium sulfate, concentrate the filtrate, use eluent (V dichloromethane) to elute through column chromatography, after elution, spin out the solvent with a rotary evaporator, and place the residue in a vacuum drying oven to dry to obtain a white solid. Yield: 75.3%. 1H NMR (400MHz, CDCl3) δ 8.34 (s, 1H), 8.07 (s, 1H), 7.93 (d, J = 8.1Hz, 1H), 7.85 (d, J = 7.7Hz, 1H), 7.45 (t, J = 8.0Hz, 1H), 4.22 (s, 2H), 3.93 (s, 3H).

[0020] (5) Synthesis of methyl 4-(2-chloroethaneylamino)benzoate (intermediate compound 6e) Take a 50ml single-mouth bottle, add 200μL of methyl p-aminobenzoate, add 267.76μL of triethylamine, add quantitative dichloromethane to dissolve it, add 298.86μL of chloroacetyl chloride, and stir at room temperature for 4 hours. Extract with dichloromethane and sodium chloride solution, dry the organic phase with anhydrous magnesium sulfate, concentrate the filtrate, use eluent (V dichloromethane) to elute through column chromatography, after elution, spin out the solvent with a rotary evaporator, and place the residue in a vacuum drying oven to dry to obtain a white solid. Yield: 75.3%. 1 H NMR (400MHz, CDCl3) δ8.38 (s, 1H), 8.05 (d, J = 8.6Hz, 2H), 7.65 (d, J = 8.6Hz, 2H), 4.21 (s, 2H), 3.91 (s, 3H).

[0021] Synthesis of target compounds 7a-7e (1) Synthesis of 2-adamantan-1-yl-N-(4-ethoxyphenyl)acetamide (Compound 7a) Take a 100ml single-mouth bottle, add 0.1g of 5-(adamantan-1-yl)-2-hydroxyphenol formaldehyde, 0.13g of 2-chloro-N-phenylacetamide, 0.1g of cesium carbonate, 0.1g of potassium iodide, 2mL of acetone and 3.5mL of NN-dimethylformamide to dissolve it at room temperature, heat the entire reaction system, connect a condenser, protect with nitrogen, and react for 6 hours. After stopping the reaction, cool to room temperature, dilute the reaction mixture with water, extract with ethyl acetate, wash with water several times, dry with anhydrous magnesium sulfate for 30 minutes, filter after drying, concentrate the filtrate, use eluent (V ethyl acetate: V petroleum ether = 1:10) to elute through 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: 60%. 1H NMR (400MHz, CDCl3) δ10.13(s,1H),9.90(s,1H),7.85(d,J=7.9Hz,2H),7.75(d,J=2.2Hz,1H),7.62(d,J=8.6Hz,1H),7.38(t ,J=7.7Hz,2H),7.15(t,J=7.4Hz,1H),6.94(d,J=8.6Hz,1H),4.69(s,2H),2.14(s,3H),1.93(s,6H),1.79(d,J=12.7Hz,6H).

[0022] (2) Synthesis of 2-(4-((3r, 5r, 7r)-adamantan-1-yl)-2-formylphenoxy)-N-(3-methoxyphenyl)acetamide (Compound 7b) Take a 100ml single-mouth bottle, add 0.1g of 5-(adamantan-1-yl)-2-hydroxyphenol formaldehyde, 0.34g of 2-chloro-N-(3-methoxyphenyl)-acetamide, 0.1g of cesium carbonate, 0.1g of potassium iodide, 2mL of acetone and 3.5mL of NN-dimethylformamide to dissolve it at room temperature, heat the entire reaction system, connect a condenser, protect with nitrogen, and react for 6 hours. After stopping the reaction, cool to room temperature, dilute the reaction mixture with water, extract with ethyl acetate, wash with water several times, dry with anhydrous magnesium sulfate for 30 minutes, filter after drying, concentrate the filtrate, use eluent (V ethyl acetate: V petroleum ether = 1:10) to elute through 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: 75%. 1 HNMR (400MHz, CDCl3) δ10.12(s,1H),9.88(s,1H),7.75(s,1H),7.61(d,J=14.0Hz,2H),6.99(dd,J=37.2,7.7Hz,3H) ,6.72(t,J=8.7Hz,3H),4.68(s,2H),3.84(s,3H),2.15(d,J=13.3Hz,3H),1.93(s,5H),1.79(q,J=12.9,12.5Hz,6H).

[0023] (3) Synthesis of methyl 2-(2-(4-((3r, 5r, 7r)-adamantan-1-yl)-2-formylphenoxy)acetylamino)benzoate (Compound 7c) Take a 100ml single-mouth bottle, add 0.1g of 5-(adamantan-1-yl)-2-hydroxyphenol formaldehyde, 0.18g of methyl 2-(2-chloroethane acylamino)benzoate, 0.1g of cesium carbonate, 0.1g of potassium iodide, 2mL of acetone and 3.5mL of NN-dimethylformamide to dissolve it, heat the whole reaction system, connect a condenser, protect with nitrogen, and react for 6 hours. After stopping the reaction, cool to room temperature, dilute the reaction mixture with water, extract with ethyl acetate, wash with water several times, dry with anhydrous magnesium sulfate for 30 minutes, filter after drying, concentrate the filtrate, use eluent (V ethyl acetate: V petroleum ether = 1:10) to elute through 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: 50%. 1 HNMR (400MHz, CDCl3) δ10.88(s,1H),10.43(s,1H),8.07(d,J=7.9Hz,1H),7.96(d,J=2.5Hz,2H),7.61(t,J=8.2Hz,2H),7 .18(t,J=7.6Hz,1H),6.99(d,J=8.7Hz,1H),4.79(s,3H),3.93(s,3H),2.13(s,3H),1.92(s,6H),1.79(d,J=10.3Hz,6H).

[0024] (4) Synthesis of methyl 3-(2-(4-((3r, 5r, 7r)-adamantan-1-yl)-2-formylphenoxy)acetylamino)benzoate (Compound 7d) Take a 100ml single-mouth bottle, add 0.1g of 5-(adamantan-1-yl)-2-hydroxyphenol formaldehyde, 0.18g of methyl 3-(2-chloroethane acylamino)benzoate, 0.1g of cesium carbonate, 0.1g of potassium iodide, 2mL of acetone and 3.5mL of NN-dimethylformamide to dissolve it, heat the whole reaction system, connect a condenser, protect with nitrogen, and react for 6 hours. After stopping the reaction, cool to room temperature, dilute the reaction mixture with water, extract with ethyl acetate, wash with water several times, dry with anhydrous magnesium sulfate for 30 minutes, filter after drying, concentrate the filtrate, use eluent (V ethyl acetate: V petroleum ether = 1:10) 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: 61%. 1HNMR (400MHz, CDCl3) δ10.88(s,1H),10.43(s,1H),8.07(d,J=7.9Hz,1H),7.96(d,J=2.5Hz,2H),7.61(t,J=8.2Hz,2H), 7.18(t,J=7.6Hz,1H),6.99(d,J=8.7Hz,1H),4.79(s,3H),3.93(s,3H),2.13(s,3H),1.92(s,6H),1.79(d,J=10.3Hz,6H).

[0025] (5) Synthesis of methyl 4-(2-(4-((3r, 5r, 7r)-adamantan-1-yl)-2-formylphenoxy)acetylamino)benzoate (Compound 7e) Take a 100ml single-mouth bottle, add 0.1g of 5-(adamantan-1-yl)-2-hydroxyphenol formaldehyde, 0.18g of methyl 4-(2-chloroethane acylamino)benzoate, 0.1g of cesium carbonate, 0.1g of potassium iodide, 2mL of acetone and 3.5mL of NN-dimethylformamide to dissolve it at room temperature, heat the entire reaction system, connect a condenser, protect with nitrogen, and react for 6 hours. After stopping the reaction, cool to room temperature, dilute the reaction mixture with water, extract with ethyl acetate, wash with water several times, dry with anhydrous magnesium sulfate for 30 minutes, filter after drying, concentrate the filtrate, use eluent (V ethyl acetate: V petroleum ether = 1:10) 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: 57%. Yield: 57%. 1H NMR (400MHz, CDCl3) δ10.29(s,1H),10.08(s,1H),8.02(s,2H),8.00(s,2H),7.97(s ,2H),7.26(s,2H),4.69(s,2H),3.91(s,3H),2.13(s,3H),1.92(s,6H),1.85(s,7H).

[0026] Example 3: In vitro cytotoxicity evaluation results 1. The present invention conducted CCK-8 experimental tests on target compounds 7a-7f, expressed them in HeLa, A549, HeGP2 and L929, and tested target compounds 7a-7f and 5-fluorouracil at a concentration of 20 μM. As can be seen from the figure, 7c containing a structure of an adamantane structure with an amine compound structure grafted at the end of the benzene ring without a substituent has the best inhibitory effect on HELA cells and HepG2 cells, with inhibition rates of 63.31% and 61.01%, respectively, and 5-fluorouracil has inhibition rates of 51.20% and 39.69% on HELA cells and HepG2 cells, respectively. 7e containing a structure of an adamantane structure with an amine compound structure grafted at the end of the benzene ring containing methyl aminobenzoate has the best inhibitory effect on A549 cells, with an inhibition rate of 66.94%, while the inhibition rate of 5-fluorouracil on A549 cells is 35.95%.

[0027] 2. The present invention calculates the IC50 values ​​of compounds 7a-7f at 40μM, 20μM, 10μM, and 5μM concentrations for HeLa, A549, and HepG2 cells, among which compounds 7c and 7e have lower IC50 values ​​for A549 cells and HepG2 cells, and 7b has lower IC50 value for HeLa cells. For amine compounds with different groups connected to the benzene ring ends at position 4, the efficacy of compounds containing methyl aminobenzoate in HELA cells and A549 cells is significantly better than that of compounds containing methoxy groups. At the same position 4, in L929 cells, the efficacy of compounds containing methyl aminobenzoate is better than that of compounds containing methoxy groups, and the efficacy of compounds containing methyl aminobenzoate is still the best.

[0028] Table 1 shows the IC50 values ​​of target compounds 7a-7f against different cells.

Claims

1. A novel small drug molecule containing an adamantane structure (1), characterized in that: The molecular chemical structure is as follows, 2. R is R1, R2, R3 are H, OCH3, CH3 in corresponding positions.

3. The adamantane structure derivative according to claim 1, wherein: Target product:

4. The adamantane structure derivative according to claim 1, wherein: Synthesis route:

5. As claimed in claim 1, the use of the adamantane structure derivative formula (1) having multiple biological activities such as antiviral, antibacterial, and antitumor.

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