A β-elemonic acid compound, its preparation method and application

Abstract

The present invention discloses a β - elemolonic acid compound, its preparation method and application, belonging to the technical field of biomedicine. The β - elemolonic acid compound of the present invention can inhibit the activity of α - glucosidase. It is based on the natural product β - elemolonic acid for structural modification and optimization to improve its physicochemical properties, enhance its pharmacological activity and improve its drug - like properties. The preparation method of the compound of the present invention has the advantages of rich raw material sources, mild reaction conditions, simple operation in the reaction process, and cheap and easily available reagents. Experiments show that the β - elemolonic acid compound of the present invention inhibits the activity of α - glucosidase significantly stronger than β - elemolonic acid and the positive control drug acarbose; at the same time, the β - elemolonic acid derivative can significantly inhibit the increase of post - prandial blood glucose value in mice and has research value for application in hypoglycemic drugs.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a β-elemonic acid compound, a preparation method thereof, and an application thereof. Background Art

[0002] α-Glucosidase is a digestive enzyme mainly present in intestinal epithelial cells and the pancreas, and can hydrolyze compounds bound by glucosidic bonds. α-Glucosidase plays an important role in human metabolism, and its activity regulation has become a new strategy for treating related diseases such as hyperglycemia. Inhibiting the activity of α-glucosidase can slow down the absorption of carbohydrates, thereby maintaining blood sugar balance, and is widely used in the treatment of metabolic diseases such as diabetes and obesity.

[0003] Currently, a variety of α-glucosidase inhibitor drugs have been developed, such as acarbose, voglibose, etc. However, long-term use of these drugs may cause some adverse reactions, such as nausea, diarrhea, etc. Therefore, the research and development of highly efficient and safe α-glucosidase inhibitors is of great significance for the treatment of metabolic diseases. The research on extracting hypoglycemic components from natural products is of great significance. On the one hand, it can provide a safer and more effective drug source, avoiding the adverse reactions that may be brought about by synthetic drugs; on the other hand, natural products are widely sourced, have complex structures, and diverse pharmacological effects, so they can provide strong support and inspiration for the research and development of new drugs. β-Elemonic acid compounds exhibit strong α-glucosidase inhibitory activity and have important application values in the medical field. Summary of the Invention

[0004] To solve the above problems, the present invention provides a β-elemonic acid compound, a preparation method thereof, and an application thereof. This compound exhibits strong α-glucosidase inhibitory activity and can be applied to the preparation of hypoglycemic drugs and pharmaceutical compositions containing it. The preparation method of this compound also has the advantages of rich raw material sources, mild reaction conditions, simple operation in the reaction process, and cheap and easily available reagents.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] One object of the present invention is to provide a β-elemonic acid compound with the following structural formula:

[0007]

[0008] The above-mentioned β-elemonic acid compound is named as:

[0009] (S)-6-methyl-2-((5R,10S,13S,14S,17S,E)-4,4,10,13,14-pentamethyl-3-oxo-2-(quinolin-2-ylmethylene)-2,3,4,5,6,7,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)hept-5-enoic acid is abbreviated as A1; the molecular formula is: C 40 H 51 NO3; the molecular weight is: 593.39.

[0010] (S)-6-methyl-2-((5R,10S,13S,14S,17S,E)-4,4,10,13,14-pentamethyl-3-oxo-2-(pyridin-2-ylmethylene)-2,3,4,5,6,7,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)hept-5-enoic acid is abbreviated as A2; the molecular formula is: C 36 H 49 NO3; the molecular weight is: 543.37.

[0011] The second object of the present invention is to provide a preparation method of the β-elemoneic acid compound, comprising the following steps:

[0012] (1) Add β-elemoneic acid and 2-quinolinecarboxaldehyde / 2-pyridinecarboxaldehyde to absolute ethanol, then add potassium hydroxide, and react at room temperature for 12-24 h. Use thin layer chromatography and 10% sulfuric acid ethanol solution to detect the reaction progress;

[0013] (2) After β-elemoneic acid completely disappears, pour the reaction solution into ice water, adjust the pH value to acidic with hydrochloric acid, extract with ethyl acetate, wash the organic layer, dry, filter by suction, and rotary evaporate the solvent under reduced pressure;

[0014] (3) Use silica gel column chromatography for separation to obtain compounds A1 and A2.

[0015] Further, the molar ratio of β-elemoneic acid, 2-quinolinecarboxaldehyde / 2-pyridinecarboxaldehyde and potassium hydroxide in step (1) is 1:1-3:1.5-3.

[0016] Further, the pH value in step (2) is less than 4; the extraction is carried out three times with 10 mL of ethyl acetate.

[0017] Further, the washing in step (2) is carried out with saturated sodium chloride solution; the drying is carried out with anhydrous sodium sulfate.

[0018] The third object of the present invention is to provide an application of the β-elemoneic acid compound in the preparation of α-glucosidase inhibitors and hypoglycemic drugs.

[0019] Further, a pharmaceutical preparation is prepared by using the β-elemoneic acid compound as an active ingredient and containing one or more pharmaceutically acceptable carrier substances and / or adjuvants.

[0020] Advantages of the present invention:

[0021] The present invention provides a β-elemoneic acid compound, which exhibits strong α-glucosidase inhibitory activity. The IC 50 values of compounds A1 and A2 are 3.68 μM and 9.00 μM respectively. The postprandial blood glucose level experiment shows that compounds A1 and A2 can significantly reduce the postprandial blood glucose level of mice, which is comparable to the positive control drug acarbose. The preparation method of the compounds of the present invention also has the advantages of rich raw material sources, mild reaction conditions, simple operation in the reaction process, and cheap and easily available reagents. Description of the drawings

[0022] Figure 1 is the structural formula of the β-elemoneic acid compound;

[0023] Figure 2 is the blood glucose measurement curve graph, including a blank control group, a compound A1 group, a compound A2 group, a positive control acarbose group, a model group, and a lead compound group (β-EA). Specific embodiments

[0024] To enable those skilled in the art to better understand the technical solution of the present invention, the present invention will be further described in detail below with reference to the embodiments.

[0025] Example 1

[0026] β-Elemoneic acid (1 mmol, 454 mg) and 2-quinolinecarboxaldehyde (3 mmol, 471 mg) / 2-pyridinecarboxaldehyde (3 mmol, 321 mg) were added to 10 mL of anhydrous ethanol, and then potassium hydroxide (3 mmol, 168 mg) was added. The reaction was carried out at room temperature for 24 h, and the reaction progress was detected by thin layer chromatography and 10% sulfuric acid ethanol solution. After β-elemoneic acid completely disappeared, the reaction solution was poured into ice water, and the pH was adjusted to 3 with 10% hydrochloric acid. Then it was extracted with ethyl acetate (3 × 10 mL). The organic layer was washed with saturated sodium chloride solution and dried with anhydrous sodium sulfate, filtered by suction, and the solvent was rotary evaporated under reduced pressure. Compounds A1 and A2 were obtained by silica gel column chromatography.

[0027] The 1H NMR data are as follows: white solid; yield: 37%; 1 1H NMR (300 MHz, CDCl3) δ 8.15 (d, J = 8.5 Hz, 1H), 8.04 (d, J = 8.7 Hz, 1H), 7.78 (t, J = 7.3 Hz, 2H), 7.56 (d, J = 7.9 Hz, 1H), 7.50 (d, J = 8.6 Hz, 2H), 5.11 (s, 1H), 3.64 (d, J = 18.6 Hz, 1H), 2.99 (d, J = 17.0 Hz, 1H), 2.40 - 2.30 (m, 2H), 2.13 - 1.95 (m, 7H), 1.83 (d, J = 12.3 Hz, 1H), 1.73 (d, J = 7.8 Hz, 1H), 1.67 (s, 3H), 1.60 (s, 2H), 1.57 (s, 3H), 1.59 - 1.53 (m, 2H), 1.49 - 1.41 (m, 4H), 1.27 (s, 3H), 1.19 (s, 3H), 1.14 (s, 3H), 0.97 (s, 3H), 0.75 (s, 3H); 13 13C NMR (126 MHz, CDCl3) δ 209.42, 182.36, 155.63, 148.07, 140.69, 135.87, 135.39, 133.70, 132.28, 131.57, 129.90 (d, J = 12.0 Hz), 127.40, 126.94, 126.65, 124.22, 123.56, 50.00, 49.87, 47.01, 45.36, 44.03, 42.68, 36.42, 32.46, 29.70, 29.55, 29.31, 28.92, 27.38, 26.99, 26.00, 25.67, 24.03, 21.85, 21.22, 20.68, 19.52, 17.61, 15.90; ESI - HRMS (m / z): calcd for C 40 H 52 NO3 + [M + H] + : 594.39417, found: 594.39422.

[0028] The 1H NMR data of A2 are as follows: white solid; yield: 42%; 1HNMR(300MHz,CDCl3)δ8.73(d,J=4.0Hz,1H),7.77 - 7.66(m,1H),7.39(d,J=8.4Hz,2H),7.17(dd,J=7.1,5.0Hz,1H),5.11(s,1H),3.48(d,J=18.2Hz,1H),2.66(d,J=17.7Hz,1H),2.33(s,1H),2.25 - 1.94(m,8H),1.81(m,2H),1.68(s,3H),1.59(s,3H),1.46 - 1.24(m,8H),1.18(s,3H),1.12(s,3H),0.94(d,J=9.9Hz,3H),0.90(s,3H),0.75(s,3H). 13 C NMR(126MHz,CDCl3)δ209.27,182.32,155.49,149.61,138.75,136.09,135.29,134.44,132.24,131.45,126.64,123.62,122.24,49.93,47.72,47.00,45.32,43.91,41.37,36.57,32.49,29.71,29.52,29.30,28.72,27.34,26.92,25.98,25.70,24.38,21.97,21.31,20.81,19.62,17.65,15.94.ESI - HRMS(m / z):calcd for C 36 H 50 NO3 + [M + H] + :544.3785,found:544.3779.

[0029] Test Example 1

[0030] 20 μM (ranging from 0.1 to 10 mM) of each sample dissolved in DMSO was added to a 96 - well plate containing 100 μL of α - glucosidase solution (pH 6.9, 0.1 U / mL, 0.1 M phosphate buffer). After mixing well, it was incubated at 25 °C for 10 min. Then, 50 μL of pNPG solution (pH 6.9, 5 mM, 0.1 M phosphate buffer) was added to each well and incubated at 25 °C for 5 min. Before and after incubation, the absorbance at 405 nm was recorded on a microplate reader. Acarbose was used as a positive control. The α - glucosidase inhibitory activity was expressed as the inhibition rate %, and the calculation method was as follows:

[0031] Inhibition rate (%)=(1 - δsample / δblank)×100%, IC 50The value was calculated by plotting the concentration-inhibition rate curve, and the IC 50 value is shown in Table 1.

[0032] Table 1 Pharmacological activity data of the target compounds

[0033] Compound <![CDATA[IC 50 Value (μM)]]> A1 3.68±0.23 A2 9.00±0.41 β-Elemoneic acid 16.2±0.62 Acarbose 342.04±2.45

[0034] Experimental Example 2

[0035] The postprandial blood glucose level was detected to explore the effects of β-elemoneic acid compounds A1 and A2 on the postprandial blood glucose level of normal mice.

[0036] Experimental materials: Male C57BL / 6 mice.

[0037] Experimental method: In this experiment, all experimental mice were fasted for 12 hours (with free access to water) before the experiment. All C57 mice were randomly divided into 5 groups, with 6 mice in each group. They were: model group (0.5% CMC-Na solution), compound A1 group (compound A1 25 mg / kg, dissolved in 0.5% CMC-Na solution), compound A2 group (compound A2 25 mg / kg, dissolved in 0.5% CMC-Na solution), lead compound group (β-EA 25 mg / kg, dissolved in 0.5% CMC-Na solution), positive control acarbose group (acarbose 25 mg / kg, dissolved in 0.5% CMC-Na solution), and blank control group (0.5% CMC-Na solution). After administration, except for the blank control group which was given 0.5% CMC-Na solution, the other groups were intragastrically administered 2.5 g / kg sucrose. At 0 min, 15 min, 30 min, 60 min, 90 min, and 120 min after administration, the postprandial blood glucose of the mice was detected from the tail using a blood glucose meter. Male C57BL / 6 mice were purchased from the Animal Experiment Center of Yanbian University.

[0038] Experimental results: The experimental results are as Figure 2 shown. In the model group, the blood glucose rose rapidly and reached the peak after oral administration of sucrose. After oral administration of sucrose in the administration groups, the level of blood glucose rise was significantly lower than that in the model group, and the blood glucose decreased steadily. The hypoglycemic ability of the compound A1 group was better than that of the acarbose group and the compound A2 group. Conclusion: Experimental Examples 1-2 fully demonstrate that compounds A1 and A2 have significant hypoglycemic ability and have research value as hypoglycemic drugs.

[0039] Experimental Example 3

[0040] Drug composition

[0041] Tablet formulation of 1000 tablets containing 100 mg of active ingredient per tablet: 100 g of compound A1, 2 g of hydroxypropyl cellulose, 10 g of wheat starch, 100 g of lactose, 3 g of magnesium stearate, 3 g of talc.

[0042] The medicine should be taken according to the doctor's advice. This pharmaceutical composition should be used starting from a small dose, and the dose should be gradually increased according to the patient's condition. The daily dose varies between 0.1 mg and 1.0 g, and can be administered once or several times. For patients who need further blood sugar control, a certain dose can be increased.

[0043] The above-described embodiments are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention should fall within the protection scope determined by the claims of the present invention.

Claims

1. A β-elemonic acid compound, characterized in that, It has the following structural formula: Compound A1: Molecular formula: C 40 H 51 NO3; Molecular weight: 593.39; Compound A2: Molecular formula: C 36 H 49 NO3; Molecular weight: 543.

37.

2. A method for preparing a β-elemolic acid compound as described in claim 1, characterized in that, It includes the following steps: (1) Add β-elemene acid and 2-quinolinecarboxaldehyde / 2-pyridinecarboxaldehyde into absolute ethanol, then add potassium hydroxide, and react at room temperature for 12 - 24 h; (2) After β-elemene acid completely disappears, pour the reaction solution into ice water, adjust the pH value to acidic with hydrochloric acid, extract with ethyl acetate, wash the organic layer, dry it, filter by suction, and rotary evaporate the solvent under reduced pressure; (3) Use silica gel column chromatography for separation to obtain compounds A1 and A2.

3. The preparation method according to claim 2, characterized in that, The molar ratio of β-elemene acid, 2-quinolinecarboxaldehyde / 2-pyridinecarboxaldehyde and potassium hydroxide described in step (1) is 1:1 - 3:1.5 - 3.

4. The preparation method according to claim 2, characterized in that, The pH value described in step (2) is less than 4; the extraction is to extract three times with 10 mL of ethyl acetate.

5. The preparation method according to claim 2, characterized in that, The washing described in step (2) is to wash with saturated brine; the drying is to dry with anhydrous sodium sulfate.

6. Use of a β-elemene acid compound as described in claim 1 in the preparation of an α-glucosidase inhibitor and an antidiabetic drug.

7. The application according to claim 6, wherein A pharmaceutical preparation is prepared with the β-elemene acid compound as described in claim 1 as the active ingredient and containing one or more pharmaceutically acceptable carrier substances and / or adjuvants.

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