A kind of ellagitannin compound and its preparation method and application

Abstract

The present application relates to a kind of tannin compounds and its preparation method and application, the tannin compound of this ellagitannin is stable, easy to store, and the IC 50 Value of 0.00089 mmol / L, indicating that it has very strong α-glucosidase inhibitory activity, and is expected to be developed as a new lead compound for the treatment of diabetes.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine, in particular to a kind of tannin class compound and its preparation method and application. BACKGROUND

[0002] Diabetes is a metabolic disease characterized by high blood sugar. High blood sugar is caused by genetic factors or environmental factors, etc. Insulin secretion defects or impaired biological effects or both. Long-term high blood sugar can cause chronic damage to various tissues of the human body, especially the eyes, kidneys, heart, blood vessels and nerves, and functional disorders, which has seriously threatened human life and health.

[0003] The therapeutic drugs for diabetes mainly include sulfonylurea drugs, biguanide hypoglycemic drugs, alpha glucosidase inhibitors, insulin sensitizers and other drugs. However, these drugs currently used have certain side effects to varying degrees. For example, acarbose, an alpha glucosidase inhibitor, has main adverse reactions such as abdominal pain, intestinal distension, diarrhea, increased anal exhaust, etc. Therefore, it is of great significance to find safe and efficient therapeutic drugs for diabetes from natural products to protect human life and health.

[0004] Tannin is a kind of complex phenolic secondary metabolites widely existing in plant bodies, which is typical of ortho-phenolic hydroxyl groups (such as o-diphenol and pyrogallol). It has strong water solubility, is unstable under light and heat, is difficult to separate and identify, and has strong biological activity. The main feature of tannin class components is that the compounds contain ellagic acid (EA) units derived from the dimerization of gallic acid. Modern pharmacological studies have shown that tannin class components have good antioxidant, anti-inflammatory, anti-tumor and other biological activities, and have potential prevention and treatment effects on chronic diseases such as cardiovascular and cerebrovascular diseases, diabetes and neuropathies. SUMMARY

[0005] The technical problem to be solved by the present application is to provide a drug for treating diabetes. The tannin class compound of the present application has very strong alpha-glucosidase inhibitory activity and can be used for treating diabetes.

[0006] The technical solution of the present application to solve the above technical problem is as follows: a tannin class compound, the structural formula of the tannin class compound is:

[0007]

[0008] The tannin class component (named 1-desgalloyleugeniin) provided by the present application has the advantages of stable properties and easy storage. The in vitro pharmacological experiment proves that the IC 50 value of the alpha-glucosidase of the tannin class component is 0.00089 mmol / L (the IC50 The results show that the strong α-glucosidase inhibitor component has very strong α-glucosidase inhibitory activity, and is expected to be used as a lead compound to develop a new diabetes treatment drug. The 1-desgalloyl eugeniin or a derivative or a pharmaceutically acceptable salt thereof can be used for a blood sugar lowering drug.

[0009] The present application provides a preparation method of the tannin component for realizing the second purpose, comprising:

[0010] The Yuanjiang acerola leaf is crushed and extracted, and then the filtrate is concentrated, water-floated, and degreased. The degreased filtrate is sequentially separated by Sephadex LH-20 gel column chromatography, MCI gel CHP 20P column chromatography, and Toyopearl HW-40F column chromatography, and the eluent is a mixture of methanol and water.

[0011] The strong α-glucosidase inhibitor component is extracted and separated from the Yuanjiang acerola leaf, the preparation method is simple and easy to operate, the leaf is used as a raw material for extraction, the resource is abundant, and the method has good potential economic benefits. The α-glucosidase inhibitor component 1-desgalloyl eugeniin has stable properties and is easy to store, has good α-glucosidase inhibitory activity, and is expected to be used as a lead compound to develop a new diabetes treatment drug.

[0012] Further, the eluent of the Sephadex LH-20 gel column chromatography, the MCI gel CHP 20P column chromatography, and the Toyopearl HW-40F column chromatography is a mixture of methanol and water with gradient concentrations, and the volume content of methanol in the mixture of methanol and water with gradient concentrations is 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% in turn.

[0013] Further, the extraction includes: extracting 2-3 times with methanol with a volume concentration of 65-75%, and each time for 6-8 days, and combining the filtrate.

[0014] The reagent used for the degreasing is petroleum ether.

[0015] The present application provides a pharmaceutically acceptable salt of the tannin component for realizing the third purpose, and the pharmaceutically acceptable salt is a salt synthesized by the tannin component and an organic acid and / or an inorganic acid.

[0016] The present application provides a pharmaceutically acceptable esterified derivative of the tannin component for realizing the fourth purpose, and the pharmaceutically acceptable esterified derivative is prepared by esterification of the tannin component.

[0017] The application provides a use of a tannin compound in the preparation of a blood sugar reducing agent.

[0018] It should be noted that the tannin compound 1-desgalloyl eugeniin or a derivative thereof or a pharmaceutically acceptable salt thereof provided by the application can be combined with excipients or carriers allowed by preparations or drugs to prepare a drug or a pharmaceutical composition for treating diabetes with α-glucosidase inhibitory activity. The drug or the pharmaceutical composition can be in the form of tablets, granules, capsules and the like; and can also adopt preparations known in the modern pharmaceutical industry such as nano, controlled release, sustained release preparations and the like.

[0019] Further, the blood sugar reducing agent comprises a pharmaceutically acceptable carrier and the tannin compound and / or the derivative of the tannin compound.

[0020] It should be noted that the derivative of the tannin compound comprises a pharmaceutically acceptable salt of the tannin compound. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 The structural formula of the tannin compound (1-desgalloyl eugeniin) of the application is shown in the following formula (I):

[0022] Figure 2 The structural formula of the tannin compound (1-desgalloyl eugeniin) of the application is shown in the following formula (I): 1 H-NMR spectrum;

[0023] Figure 3 The structural formula of the tannin compound (1-desgalloyl eugeniin) of the application is shown in the following formula (I): 13 C-NMR spectrum. DETAILED DESCRIPTION

[0024] The principles and characteristics of the application are described below, and the examples are only used to explain the application and are not used to limit the scope of the application. If the specific technology or condition is not indicated in the examples, the technology or condition is performed according to the technology or condition described in the literature in the art or according to the product instruction. If the reagent or instrument is not indicated by the manufacturer, it is a conventional product that can be commercially available through a regular channel.

[0025] EMBODIMENT

[0026] A tannin compound (1-desgalloyl eugeniin) and a preparation method thereof, comprising:

[0027] (1) Dry Yuanjiang cone leaf 5.5 kg was crushed and soaked in 40 L of 70% methanol for extraction for 3 times, each time for 7 days. The filtrate was combined and concentrated under reduced pressure to obtain extract. The extract was defatted with 1.5 L of petroleum ether for 3 times. The water solution after removing petroleum ether was concentrated under reduced pressure to remove petroleum ether;

[0028] (2) The water solution after removing petroleum ether was separated by Sephadex LH-20 gel (10 cm x 50 cm) column chromatography. Gradient elution was performed with 2 L of methanol and water mixed solution with gradient concentration (the volume fraction of methanol was 0%, 20%, 40%, 60%, 80%, and 100% successively) successively. TLC was used to monitor and combine the same components to obtain 7 fractions (Fr. 1-7);

[0029] (3) Fr. 4 (227.0 g) was separated by MCI gel CHP 20P (7.5 cm x 29.5 cm) column chromatography. Gradient elution was performed with 0.8 L of methanol and water mixed solution with gradient concentration (the volume fraction of methanol was 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% successively) successively. TLC was used to monitor and combine the same components to obtain 11 fractions (Fr. 4.1-4.11);

[0030] (4) Fr. 4.5 (430 mg) was separated by Toyopearl HW-40F (3.5 cm x 32.0 cm) column chromatography. Gradient elution was performed with 0.5 L of methanol and water mixed solution with gradient concentration (the volume fraction of methanol was 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% successively) successively. 44 mg of tannin compound (1-desgalloyl eugeniin) was obtained.

[0031] Identification of tannin compound (1-desgalloyl eugeniin) of the example:

[0032] Figure 2 is the tannin compound (1-desgalloyl eugeniin) 1 H-NMR spectrum; Figure 3 is the tannin compound (1-desgalloyl eugeniin) 13 C-NMR spectrum. The tannin compound (1-desgalloyl eugeniin) is white amorphous powder, has bright spots under 254 nm ultraviolet light, and shows brown spots after reacting with 1% FeCl3-EtOH color reagent. 1H-NMR (500 MHz, acetone-d6), 13 C-NMR (125 MHz, acetone-d6) data are shown in Table 1 below.

[0033] Table 1 NMR data of the ellagitannin compound (1-desgalloyl eugeniin) (in acetone-d6)

[0034]

[0035]

[0036] According to the comprehensive analysis of the above spectrum data, the structure of the ellagitannin compound (1-desgalloyl eugeniin) is deduced as shown in Figure 1 .

[0037] Activity detection of the ellagitannin compound (1-desgalloyl eugeniin):

[0038] Prepare 20x PBS phosphate buffer saline with a concentration of 50 mM and a pH value of 6.8, use 1 mmol / L of p-nitrophenyl-α-D-glucopyranoside PNPG as the substrate, and prepare 10 U / mg of purchased α-glucosidase into an enzyme solution with an enzyme activity of 0.5 U / mL for standby, and acarbose as a positive control. Add 50 μL of PBS phosphate buffer to a 96-well plate, add 40 μL of different concentrations of samples (1-desgalloyl eugeniin) and PBS to the enzyme reaction group and the blank group respectively, add 10 μL of α-glucosidase, mix well, incubate at 37°C for 5 min, then add 20 μL of 1 mmol / L PNPG solution, and react in a 37°C incubator for 30 min. Then, use an enzyme marker to measure the absorbance value at 405 nm wavelength.

[0039] According to the formula: α-glucosidase inhibition rate = [1-(A 反应组 -A 反应对照组 ) / (A 空白组 -A 空白对照组 )]x 100%, calculate the enzyme activity inhibition rate. The above experiment is repeated 3 times in parallel, and the average value is calculated to calculate the IC 50 value.

[0040] Activity results:

[0041] The IC 50 of the positive drug acarbose is 0.0019 mmol / mL, and the IC 50The compound has 0.00089mmol / L of α-glucosidase inhibitory activity, can be used for preparing hypoglycemic drugs, opens up a new way for preparing hypoglycemic drugs and a new use of Yuanjiang cone plants, and has significant economic and social benefits.

[0042] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "an example", "a specific example", or "some examples" and the like means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples. Furthermore, different embodiments or examples described in the present specification and the features of different embodiments or examples can be combined and combined by those skilled in the art without contradiction.

[0043] Although the embodiments of the present application have been shown and described above, it is understood that the above-described embodiments are exemplary and are not to be construed as limiting the present application, and those skilled in the art can make changes, modifications, replacements and variations to the above-described embodiments within the scope of the present application.

Claims

1. The application of ellagitannins in the preparation of hypoglycemic drugs, characterized in that, The structural formula of the ellagitannin compound is: 。 2. The application of the ellagitannins according to claim 1 in the preparation of hypoglycemic drugs, characterized in that, The hypoglycemic drug includes a pharmaceutically acceptable carrier and the ellagitannin compound.

3. The application of the ellagitannins according to claim 1 in the preparation of hypoglycemic drugs, characterized in that, The preparation method of the ellagitannin compounds includes the following steps: After crushing and extracting the leaves of *Cephalotaxus fortunei*, the filtrate was concentrated, floated in water, and defatted. The defatted filtrate was then separated sequentially by Sephadex LH-20 gel column chromatography, MCI gel CHP 20P column chromatography, and Toyopearl HW-40F column chromatography, with the eluent being a mixture of methanol and water.

4. The application of the ellagitannin compounds according to claim 3 in the preparation of hypoglycemic drugs, wherein the eluents for the Sephadex LH-20 gel column chromatography, the MCI gel CHP 20P column chromatography, and the Toyopearl HW-40F column chromatography are mixtures of methanol and water with gradient concentrations, wherein the methanol volume content of the mixtures of methanol and water with gradient concentrations is successively 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100%.

5. The application of the ellagitannins according to claim 3 in the preparation of hypoglycemic drugs, wherein the extraction comprises: Extract with methanol at a volume concentration of 65-75% 2-3 times, each extraction lasting 6-8 days, and combine the filtrates; The reagent used for degreasing is petroleum ether.

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