Fluorine-containing N-aryl valine derivative and application thereof

Abstract

The invention discloses a fluorine-containing N-aryl valine derivative with the following structural formula, the fluorine-containing N-aryl valine derivative is synthesized by taking p-hydroxybenzoic acid as a raw material through amidation, etherification, halogenation, hydrolysis and other steps, and the fluorine-containing N-aryl valine derivative can enhance uptake and consumption of glucose by HepG2 cells, effectively improve cell insulin resistance, improve the activity of glucose in the cells and improve the activity of glucose in the cells. Good in-vitro hypoglycemic activity is achieved, and a new medicine is provided for preventing or treating glucose metabolism disorder; # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of medical technology and mainly relates to a fluorine-containing N-aryl valine derivative and application thereof. Background Art

[0002] Organic fluorine compounds account for 20% to 25% of the drugs currently on the market and 30% to 40% of the pesticides on the market, playing an important role in pharmaceutical chemistry. The introduction of fluorine atoms or fluorine-containing groups will change the physical and chemical properties of the compounds, so fluorine-containing compounds are widely used in many fields such as medicine and pesticides. For example, the fluorine-containing drugs on the market include the cholesterol-lowering drug atorvastatin, the anti-tumor drug vinflunine, and the drug Iressa for the treatment of lung cancer. Fluorination will change the binding affinity between small molecules and proteins, thereby affecting the activity and drugability of the compounds. Fluorine-containing drugs have anti-caries, antibacterial, anti-inflammatory, bone protection, lowering blood cholesterol levels, and anti-tumor functions. For example, fluorine-containing amino acids have been used as antiviral and anti-tumor agents, and the combination of fluorine-containing amino acids with proteins can enhance the activity of proteins. The fluorine-containing drug fluvastatin sodium is a commonly used lipid-lowering drug with good lipid-lowering effects. It is a methylglutaryl coenzyme A reductase inhibitor and mainly works in the liver. Fluorinated amino acids and their derivatives are a class of compounds with special biological activity, have potential medicinal value, and are expected to become an important member of synthetic drugs. However, there are few reports on the synthesis and functional application of fluorinated N-aryl valine derivatives. Summary of the invention

[0003] The present invention provides a fluorine-containing N-aryl valine derivative, the chemical structure of which is as follows:

[0004]

[0005] The fluorinated N-aryl valine derivative of the present invention is synthesized from p-hydroxybenzoic acid as a raw material through amidation, etherification, halogenation, and hydrolysis steps, and the specific steps are as follows:

[0006]

[0007] (1) Amidation reaction

[0008] Add p-hydroxybenzoic acid, L-valine hydrochloride, a condensing agent, dichloromethane and a base into a three-necked flask, and stir to react at room temperature. After the reaction is completed, pour the reaction product into a separating funnel, add distilled water for extraction, separate the organic phase, dry it with anhydrous sodium sulfate, filter it, evaporate the filtrate to dryness, and perform silica gel chromatography to obtain intermediate 1;

[0009]

[0010] The condensing agent is selected from DCC, EDC, and Carter's condensing agent (BOP), and the base is selected from triethylamine, pyridine, and diisopropylethylamine (DIEPA);

[0011] (2) Etherification reaction

[0012] Triethylene glycol and pyridine were added to dichloromethane, cooled to 0°C, p-toluenesulfonyl chloride was added, and after the reaction was completed by stirring at room temperature, the reaction solution was washed with hydrochloric acid until neutral, and then washed with distilled water, the organic phase was separated, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography to obtain intermediate 2';

[0013]

[0014] Dissolve intermediate 1, intermediate 2' and triphenylphosphine in dichloromethane, cool to 0°C, add diethyl azodicarboxylate dropwise under stirring, and after the reaction is completed, wash with distilled water, separate the organic phase, dry over anhydrous sodium sulfate, filter, concentrate the filtrate and purify by silica gel column chromatography to obtain intermediate 2;

[0015]

[0016] (3) Halogenation reaction

[0017] The intermediate 2, potassium fluoride, and cryptand K222 (4,7,13,16,21,24-hexaoxo-1,10-diazabicyclo[8.8.8]hexacosane) were dissolved in anhydrous acetonitrile, and the mixture was refluxed until the raw material disappeared, and the solvent was removed by rotary evaporation. The residue was separated and purified by silica gel column chromatography to obtain the intermediate 3;

[0018]

[0019] (4) Hydrolysis reaction

[0020] The intermediate compound 3 was dissolved in alcohol, and then a 1M alkali solution was added, and the mixture was stirred at room temperature until the starting material disappeared. After the reaction, the pH was adjusted to neutral with 1M hydrochloric acid, and the mixture was extracted with ethyl acetate. The organic phase was separated, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and purified by silica gel column chromatography to obtain the final product 4 (fluorinated N-aryl valine derivative);

[0021] The alcohol is methanol or ethanol, and the base is sodium hydroxide, potassium hydroxide or sodium carbonate;

[0022]

[0023] Another object of the present invention is to use the fluorine-containing N-aryl valine derivatives in the preparation of drugs for preventing or treating glucose metabolism disorders and dyslipidemia.

[0024] The present invention uses the above-mentioned fluorinated N-aryl valine derivatives in the preparation of drugs for preventing or treating glucose metabolism disorders and dyslipidemia. The ingredients (or active ingredients) of the drugs are the above-mentioned glycosylated ganoderic acid, and one or more excipients acceptable in product production can also be added to improve the absorption effect of the active ingredients or facilitate use, such as making them into capsules or pills, powders, tablets, granules, oral liquids and injections, etc., that is, making them into suitable dosage forms for use.

[0025] The fluorinated N-aryl valine derivatives of the present invention can enhance the uptake and consumption of glucose by HepG2 cells, effectively improve cellular insulin resistance, have good in vitro hypoglycemic activity, and provide new drugs for preventing or treating glucose metabolism disorders; and the raw materials for preparing the compounds of the present invention are readily available, the conditions are mild, the method is simple and the operation is convenient, and the compounds are suitable for industrial production and market promotion applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The results show that fluorinated N-aryl valine derivatives can improve insulin resistance in HepG2 cells. DETAILED DESCRIPTION

[0027] The method of the present invention is further described in detail below through the accompanying drawings and examples, but the protection scope of the present invention is not limited to the above contents. Unless otherwise specified, the reagents in the examples are conventional reagents or reagents prepared according to conventional methods.

[0028] Example 1: Preparation of Fluorinated N-Aryl Valine Derivatives

[0029] (1) Amidation reaction

[0030] p-Hydroxybenzoic acid (2.28 g, 20 mmol), L-valine hydrochloride (3.36 g, 20 mmol), BOP (8.84 g, 20 mmol), dichloromethane (60 mL), and DIEPA (9.3 mL, 60 mmol) were added to a three-necked flask, and the mixture was stirred at room temperature for 20 h. After the reaction, the reaction product was poured into a separatory funnel, and distilled water was added in a volume ratio of 1:1 for extraction. The organic phase was separated, and the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated to dryness and then subjected to silica gel column chromatography (ethyl acetate / n-hexane = 1:5) to obtain intermediate 1 (1.96 g) with a yield of 39.0%;

[0031] Intermediate 1: 1 H NMR (400 MHz, CDCl 3)δ7.79-7.77(m,2H),7.35-7.33(m,2H),4.03-4.00(t,1H,J=6.3Hz),3.64-3.69(t,3H,J=6.3Hz),2.47(s,1H),1.33-1.26(m,6H)ppm.

[0032] through 1 H-NMR determination confirmed the structure of the compound as

[0033]

[0034] (2) Etherification reaction

[0035] Triethylene glycol (4 mL, 30 mmol) and pyridine (15 mL) were dissolved in 60 mL of dichloromethane, cooled to 0°C, p-toluenesulfonyl chloride (5.7 g, 30 mmol) was added, and the reaction was stirred at room temperature for 24 h. After the reaction, the reaction solution was washed with 1 M hydrochloric acid until neutral, and then washed with distilled water. The organic phase was separated, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography to obtain intermediate 2' (109 mg) with a yield of 20%.

[0036] Intermediate 2': 1 H NMR (400 MHz, CDCl 3 ): δ7.80-7.76(m,2H),7.34-7.32(m,2H),4.16-4.14(m,2H),3.70-3.67(m,4H),3.59-3.58(m,4H),3.57-3.54(m,2H),2.43(s,3H)ppm.

[0037] through 1 H-NMR determination confirmed the structure of the compound as

[0038]

[0039] Intermediate 1 (697 mg, 2.78 mmol), intermediate 2' (845 mg, 2.78 mmol), and triphenylphosphine (728 mg, 2.78 mmol) were dissolved in dichloromethane, cooled to 0°C, and diethyl azodicarboxylate (0.5 mL, 2.78 mmol) was added dropwise with stirring. After reacting for 6 h, the mixture was washed with distilled water, and the organic phase was separated. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated by rotary evaporation and purified by silica gel column chromatography to obtain intermediate 2, 977 mg, with a yield of 64%.

[0040] Intermediate 2: 1 H NMR (400 MHz, CDCl 3): δ7.79-7.75(m,4H),7.33-7.31(m,2H),6.94-6.92(m,2H),6.56(d,1H,J=8.8Hz),4.77-4.73(m,1H),4.14(t,4H,J= 4.8Hz),3.84-3.82(m,2H),3.75(s,3H),3.69-3.60(m,6H),2.42(s,3H),2.29-2.21(m,1H),0.98(t,6H,J=7.2Hz)ppm. 13 C NMR (100 MHz, CDCl 3 ): δ172.8,166.7,161.5,144.8,132.8,132.1,132.0,131.9,129.8,126.5, 114.3,70.7,69.5,69.2,68.7,67.5,57.3,52.2,31.6,21.6,19.0,17.9ppm.

[0041] through 1 H-NMR, 13 C-NMR determination confirmed the structure of the compound as

[0042]

[0043] (3) Halogenation reaction

[0044] Intermediate 2 (173 mg, 0.31 mmol), KF (27 mg, 0.46 mmol) and K222 (174 mg, 0.46 mmol) were dissolved in anhydrous acetonitrile and refluxed until the starting material disappeared. The solvent was removed by rotary evaporation and the residue was separated and purified by silica gel column chromatography to obtain intermediate 3, 64 mg, with a yield of 53%.

[0045] Intermediate 3: 1 H NMR (400 MHz, CDCl 3 ): δ7.78-7.75(m,2H),6.96-6.94(m,2H),6.52(d,1H,J=6.4Hz),4.79-4.75(m,1H),4.63(t,1H,J=4.2Hz),4.51(t ,1H,J=4.2Hz),4.20-4.17(m,2H),3.90-3.88(m,2H),3.80-3.71(m,6H),2.30-2.22(m,1H),1.01-0.97(m,6H)ppm. 13 C NMR (100 MHz, CDCl 3): δ172.8,166.7,161.6,128.8,126.5,114.4,100.0,84.0,82.3,70.9,70.6,70.4,69.6,67.6,57.3,52.2,31.7,19.0,18.0ppm.

[0046] through 1 H-NMR, 13 C-NMR determination confirmed the structure of the compound as

[0047]

[0048] (4) Hydrolysis reaction

[0049] The intermediate compound 3 (63 mg, 0.16 mmol) was dissolved in MeOH (2 mL), and NaOH (1 M, 20 mL) was added to the reaction solution. The mixture was stirred at room temperature until the starting material disappeared. After the reaction, the pH was adjusted to neutral with 1 M hydrochloric acid, and the mixture was extracted with ethyl acetate. The organic phase was separated and dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography to obtain the final product 4, i.e., a fluorinated N-aryl valine derivative, 39 mg, with a yield of 66%.

[0050] Product 4: 1 H NMR (400 MHz, CDCl 3 ): δ7.76-7.74(m,2H),6.94-6.92(m,2H),6.65(d,1H,J=6.4Hz),4.72-4.69(m,1H),4.62(t,1H,J=4.0Hz),4.50(t,1 H,J=4.0Hz),4.18-4.16(m,2H),3.89-3.87(m,2H),3.80-3.70(m,6H),2.36-2.28(m,1H),1.02(t,6H,J=7.2Hz)ppm. 13 C NMR (100 MHz, CDCl 3 ): δ167.4,161.7,132.2,132.1,129.1,128.9,128.5,128.7,128.5,126.3,114.5,83 .9,82.3,70.9,70.8,70.5,70.3,69.6,67.6,31.2,29.7,19.1,17.9ppm.HRMS:calcd for, found 372.1820.

[0051] through 1 H-NMR, 13 C-NMR and HRMS measurements confirmed the structure of the compound to be

[0052]

[0053] Example 2: In vitro hypoglycemic activity test of fluorinated N-aryl valine derivatives

[0054] Dexamethasone combined with insulin was used to induce HepG2 cells to establish an insulin resistance model, and the effects of fluorinated N-aryl valine derivatives on cellular glucose metabolism were studied.

[0055] HepG2 cells in logarithmic growth phase were taken at 2×10 4 Each well was inoculated in a 96-well plate and incubated at 37°C with 5% CO 2 After culturing for 12 h, the cells were treated with complete medium containing dexamethasone and insulin. A blank control group was added with complete medium, a model group was added with 0.001 μmol / L insulin and 10 μmol / L dexamethasone, and a drug-treated group was added with 0.001 μmol / L insulin, 10 μmol / L dexamethasone, and 5, 10, and 20 μmol / L fluorinated N-aryl valine derivatives. Three replicate wells were set up in each group. After culturing for 48 h, the glucose concentration in the supernatant of HepG2 cells was detected according to the instructions of the glucose detection kit, and the glucose consumption was calculated.

[0056] The results are as follows Figure 1 As shown in the figure, compared with the blank group, the glucose consumption of the model group was significantly reduced by dexamethasone combined with insulin modeling (P<0.001). Compared with the model group, the drug group significantly promoted the glucose consumption of HepG2 cells at a concentration of 20 μmol / L (P<0.05). The above results show that fluorinated N-aryl valine derivatives can enhance the uptake and consumption of glucose by cells and have good in vitro hypoglycemic activity.

Claims

1. The fluorinated N-aryl valine derivative has the structural formula shown below:

2. Use of the fluorinated N-aryl valine derivative according to claim 1 in the preparation of a drug for preventing and / or treating diabetes.

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