Perillyl alcohol derivative and application thereof

Abstract

The invention discloses a perillyl alcohol derivative with a structural formula shown in the following formula, which is prepared by the following steps: by taking perillyl alcohol as a raw material, brominating the perillyl alcohol by using carbon tetrabromide, then etherifying with a fluorine-containing benzene ring and a valine group, and finally carrying out hydrolysis reaction to obtain the perillyl alcohol derivative. The perillyl alcohol derivative prepared by the method can enhance uptake and consumption of glucose by HepG2 cells and effectively improve cell insulin resistance, has good in-vitro hypoglycemic activity, and provides a new medicine for preventing and treating glucose metabolism disorder; # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of chemical medicines, and in particular relates to a perillyl alcohol derivative and a salt thereof capable of improving insulin resistance and application thereof. Background Art

[0002] Perillyl alcohol is a natural product of monoterpenes, which is widely found in various plants such as perilla leaves and citrus. Perillyl alcohol is stable in nature, not easy to volatilize, and is mostly obtained by chemical or biological transformation. Perillyl alcohol has good anti-cancer effects, low toxicity, and is not easy to develop drug resistance. Perillyl alcohol is effective in the treatment of diseases such as glioblastoma, skin cancer, pancreatic cancer, lung cancer, liver cancer, breast cancer, prostate cancer, and lymphoma. Clinical data show that nasal targeted administration of perillyl alcohol can quickly reach the central nervous system through the blood-brain barrier and reduce side effects, but oral administration of large doses of perillyl alcohol can cause gastrointestinal discomfort, vomiting or diarrhea in patients. Perillyl alcohol is widely used in the fields of food, health products, and flavors and fragrances, but there are no reports on the use of perillyl alcohol in the prevention and treatment of diabetes. At present, research reports on perillyl alcohol derivatives mainly focus on the modification and transformation of hydroxyl groups to form esters or glycosides, and there are few reports on the formation of ether bonds. Summary of the invention

[0003] The invention provides a perillyl alcohol derivative. Perillyl alcohol is used as a raw material, and its structure is modified to introduce a fluorine-containing benzene ring and a valine group into the structure of perillyl alcohol. It is found that the compound has the ability to improve insulin resistance. The invention is of great significance to the development of hypoglycemic drugs.

[0004] The structure of the perillyl alcohol derivative of the present invention is as follows:

[0005]

[0006] The preparation method of the perillyl alcohol derivative of the present invention is as follows:

[0007]

[0008] (1) Halogenation reaction

[0009] L-perillyl alcohol, triphenylphosphine and anhydrous benzene were added to a three-necked flask, and carbon tetrabromide was added under a nitrogen atmosphere. The mixture was stirred at room temperature for reaction. After the reaction was complete, the oxidized triphenylphosphine was removed by filtration, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to obtain compound 1;

[0010]

[0011] (2) Condensation reaction

[0012] 2-Fluoro-4-hydroxybenzoic acid, L-valine methyl ester, Carter condensation agent (BOP), and diisopropylethylamine were dissolved in dichloromethane, stirred at room temperature overnight, and the reaction was followed by thin layer chromatography. After the reaction was completed, the mixture was washed three times with a saturated sodium chloride solution, and the dichloromethane layer was dried over anhydrous sodium sulfate. The solvent was dried by spin drying to obtain a crude product, which was separated and purified by silica gel column chromatography to obtain an intermediate compound 2;

[0013]

[0014] (3) Etherification reaction

[0015] Dissolve 4-hydroxybenzoic acid methyl ester-L-valine (compound 2) and cesium carbonate in dimethylformamide, stir at room temperature, then add bromoperillyl alcohol (compound 1) to continue the reaction. After the reaction is completed, remove the solvent by rotary evaporation, add dichloromethane and distilled water for extraction, collect the organic phase, dry it with anhydrous sodium sulfate, rotary evaporation, and separate it by silica gel column chromatography to obtain the intermediate compound 3;

[0016]

[0017] (4) Hydrolysis reaction

[0018] Compound 3 was dissolved in a 95% ethanol solution containing 0.2 mol / L NaOH, and stirred at room temperature overnight. After the reaction was completed, the ethanol was dried by spin drying, and ethyl acetate was added for extraction. The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was spin dried to obtain valine perilla ether 4.

[0019]

[0020] Another object of the present invention is to use the perillyl alcohol derivatives in the preparation of drugs for preventing and / or treating insulin resistance.

[0021] Another object of the present invention is to use the perillyl alcohol derivatives in the preparation of drugs for preventing and / or treating diabetes.

[0022] The drug described in the present invention, the drug ingredient (or active ingredient) is the above-mentioned perillyl alcohol derivative, and one or more excipients acceptable in product production can also be added to improve the absorption effect of the active ingredient or facilitate use, such as making it into capsules or pills, powders, tablets, granules, oral liquids and injections, etc., that is, making it into a suitable dosage form.

[0023] The perillyl alcohol derivatives of the present invention effectively improve the insulin resistance of HepG2 cells, promote the uptake of glucose by cells, and provide new ideas for drugs for preventing or treating diabetes. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1The results show the effects of perillyl alcohol derivatives on glucose consumption in IR-HepG2 cells. DETAILED DESCRIPTION

[0025] 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.

[0026] Example 1: Synthesis of Intermediate 1

[0027] L-perillyl alcohol (0.15 mL, 1.0 mmol), triphenylphosphine (524 mg, 2.0 mmol) and 8 mL of anhydrous benzene were added into a 50 mL three-necked flask, and carbon tetrabromide (663 mg, 2.0 mmol) was added under a nitrogen atmosphere. The reaction was stirred at room temperature and monitored by TLC spot plate. Solids precipitated after about 3 hours. After the raw materials reacted completely, the reaction was stopped, the reaction product was filtered to remove oxidized triphenylphosphine, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (n-hexane) to obtain the target compound 1, 380 mg, with a yield of 88.4%.

[0028] NMR data: 1 H NMR (400MHz, CDCl3) δ5.90 (s, 1H), 4.73-4.71 (d, 2H, J = 8.0Hz), 3.95 (s, 2H) ,2.23-2.14(m,4H),2.00-1.86(m,2H),1.73(s,3H),1.56-1.46(m,1H)ppm.

[0029] through 1 H-NMR determination confirmed the structure of compound 1:

[0030]

[0031] Example 2: Synthesis of Intermediate 2

[0032] 2-Fluoro-4-hydroxybenzoic acid, L-valine methyl ester, BOP and diisopropylethylamine were dissolved in dichloromethane, stirred at room temperature overnight, and the reaction was followed by thin layer chromatography. After the reaction was completed, the mixture was washed three times with a saturated sodium chloride solution, and the dichloromethane layer was dried over anhydrous sodium sulfate. The solvent was dried by spin drying to obtain a crude product, which was separated and purified by silica gel column chromatography to obtain an intermediate compound 2;

[0033] NMR data: 1H NMR (400MHz, CDCl3) δ7.80-7.85(m,1H),7.21-7.26(m,1H),6.69-6.72(m,1H),6.61-6.64(m,1H),4. 69-4.72(m,2H,J=8.0Hz),3.77(s,3H),2.24-2.32(m,4H),1.25-1.28(m,2H),0.99-1.03(m,6H)ppm.

[0034] through 1 H-NMR determination confirmed the structure of compound 2:

[0035]

[0036] Example 3: Synthesis of Intermediate 3

[0037] Compound 2 (135 mg, 0.5 mmol) and CsCO3 (212 mg, 0.65 mmol) were dissolved in DMF and stirred at room temperature. Intermediate 1 (215 mg, 1.0 mmol) was added dropwise to the reaction solution and continued to stir at room temperature. After the reaction was completed, the solvent was removed by rotary evaporation, and the mixture was extracted with dichloromethane and distilled water. The organic phase was collected and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and chromatographed on a silica gel column (dichloromethane: methanol = 95:5) to obtain intermediate compound 3.

[0038] NMR data: 1 H NMR (400MHz, CDCl3) δ7.98-8.03(s,1H),7.08-7.13(m,1H),6.77-6.80(m,1H),6.63-6.67(m,1H),5.85(s,1H),4.72-4.78(m ,3H),4.43(s,2H),3.76(s,3H),2.15-2.31(m,5H),1.86-2.04(m,2H),1.74(s,3H),1.47-1.66(m,2H),0.98-1.02(m,6H)ppm. 13 CNMR (400MHz, CDCl3) δ172.4,163.1,163.0,160.5,149.4,133.1,132.5,126 .1,113.0,112.8,111.4,72.8,57.6,52.1,40.8,31.3,30.4,27.2,26.2ppm.

[0039] through 1 H-NMR and 13 C-NMR determination confirmed the structure of compound 3:

[0040]

[0041] Example 4: Synthesis of Product 4

[0042] Compound 3 was dissolved in a 95% ethanol solution containing 0.2 mol / L NaOH and stirred at room temperature overnight. After the reaction was completed, the ethanol was dried and extracted with ethyl acetate. The ethyl acetate layer was dried over anhydrous sodium sulfate and filtered. The filtrate was dried to obtain perillyl alcohol derivative 4.

[0043] NMR data: 1 H NMR(400MHz, CDCl3)δ8.87(s,1H),7.96-8.00(m,1H),7.16-7.22(m,1H),6.64-6.78(m,1H),6.60-6.61(m,1H),5.83(s,1H),4.70-4.7 7(m,3H),4.40(s,1H),2.29-2.37(m,1H),2.13-2.20(m,4H),1.84-2.03(m,1H),1.73(s,3H),1.45-1.55(s,1H),0.99-1.04(m,6H)ppm. 13 C NMR (400MHz, CDCl3) δ175.6,163.6,163.2,163.1,163.0,160.5,149.3,133.1,133.0,132.4,126.0,11 2.6,112.5,111.3,108.8,102.4,102.1,72.7,57.6,40.7,31.0,30.3,27.1,26.1,20.7,19.0,17.6ppm.

[0044] through 1 H-NMR and 13 C-NMR determination confirmed the structure of compound 4:

[0045]

[0046] Example 5: Effect of perillyl alcohol derivatives on improving insulin resistance in HepG2 cells

[0047] HepG2 cells in the logarithmic growth phase were obtained, digested with trypsin, and added with complete culture medium to adjust the concentration to 2×10 5 The cell suspension of 100 μL / mL was inoculated into a 96-well plate, with 100 μL per well. After culturing for 12 h at 37°C and 5% CO2, the modeling reagents insulin (Ins) and dexamethasone (Dex) were added to induce the IR-HepG2 cell model.

[0048] The cells were divided into 6 groups: blank group (complete culture medium), model group (10 -3 μmol / L insulin + 10μmol / L dexamethasone), and 1 group (10 -3 μmol / L insulin + 10μmol / L dexamethasone + 2.5μmol / L perillyl alcohol derivative 4), two groups (10 -3 μmol / L insulin + 10μmol / L dexamethasone + 5μmol / L perillyl alcohol derivative 4), 3 groups (10 -3 μmol / L insulin + 10μmol / L dexamethasone + 10μmol / L perillyl alcohol derivative 4), 4 groups (10 -3 μmol / L insulin + 10μmol / L dexamethasone + 20μmol / L perillyl alcohol derivative 4), treated according to experimental groups, cultured for 48h, the culture supernatant was aspirated from each well, and the cellular glucose consumption was determined using a glucose kit.

[0049] The results are as follows Figure 1 As shown in the results, the glucose consumption of IR-HepG2 cells in the model group was significantly reduced (p < 0.01), and after being treated with different concentrations of perillyl alcohol derivatives, the glucose consumption of IR-HepG2 cells was significantly increased (p < 0.001). When cells become insulin resistant, the sensitivity of cells to insulin decreases, and insulin signal transduction is restricted, resulting in a decrease in the cell's glucose uptake. Perillyl alcohol derivatives effectively improve the insulin resistance of HepG2 cells and promote the cell's glucose uptake.

Claims

1. The structural formula of the perillyl alcohol derivative is as follows:

2. The sodium salt or potassium salt of the perillyl alcohol derivative according to claim 1.

3. Use of the perillyl alcohol derivative and salt thereof according to claim 1 or 2 in the preparation of a medicament for preventing and / or treating insulin resistance.

4. Use of the perillyl alcohol derivative and salt thereof according to claim 1 or 2 in the preparation of a drug for preventing and / or treating diabetes.

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