Phenolic compound with effect of accelerating alcohol metabolism and preparation method thereof

Abstract

The invention provides a phenolic compound with an effect of accelerating alcohol metabolism and a preparation method of the phenolic compound. The structural formula of the polyphenol compound 1-(2-hydroxyl, 4-O-glucosylphenyl)-2-(4-hydroxyphenyl) ethanone is as shown in formula (I), and the polyphenol compound 1-(2-hydroxyl, 4-O-glucosylphenyl)-2-(4-hydroxyphenyl) ethanone is separated from radix puerariae. In-vitro pharmacological experiments prove that the compound 1-(2-hydroxyl, 4-O-glucosylphenyl)-2-(4-hydroxyphenyl) ethanone has good acetylcholin esterase inhibitory activity, and the EC50 value of the compound 1-(2-hydroxyl, 4-O-glucosylphenyl)-2-(4-hydroxyphenyl) ethanone is 1.41 + / -0.12 mM, which indicates that the compound 1-(2-hydroxyl, 4-O-glucosylphenyl)-2-( The polyphenol compound 1-(2-hydroxyl, 4-O-glucosylphenyl)-2-(4-hydroxyphenyl) ethanone is expected to be used as a lead compound to develop a novel alcohol effect dispelling and liver protecting medicine.

Description

Technical Field

[0001] This invention belongs to the field of natural medicine technology, specifically relating to a novel alkaloid compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone, its preparation method, and its application in the preparation of alcohol dehydrogenase activator drugs. Background Technology

[0002] In recent years, the widespread availability of cheap alcohol has contributed to dangerous alcohol consumption, a phenomenon fueled by commercial interests, leading to a continuous increase in the number and scale of alcohol consumers. Alcohol is classified as a Group 1 carcinogen by the International Agency for Research on Cancer (IARC) of the World Health Organization, and the number of cancers is expected to rise further with increasing alcohol consumption. After ingestion, alcohol is primarily metabolized in the liver, where it is produced by alcohol dehydrogenase (ADH) to acetaldehyde. This acetaldehyde then undergoes further reactions to produce acetic acid, which is ultimately broken down into water and carbon dioxide through a series of complex oxidative respiration reactions, releasing energy. Alcohol dehydrogenase is crucial to the alcohol metabolism process, and its activity directly affects the rate of alcohol metabolism.

[0003] Kudzu is a vine belonging to the genus *Pueraria* of the legume family, including three species: *Pueraria lobata*, *Pueraria lobata*, and *Pueraria lobata* var. *pinnata*. Kudzu is rich in isoflavones and other active substances, and has been reported to have effects in relieving hangovers and protecting the liver. Currently, there are no research reports on 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone, a component in kudzu that has alcohol dehydrogenase activating activity. Summary of the Invention

[0004] The purpose of this invention is to provide a novel alkaloid component, 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone, a method for its preparation, and its application in the preparation of a drug with alcohol dehydrogenase activating activity.

[0005] The novel alkaloid component 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone and its esterified derivatives described in this invention have the structure represented by formula (Ⅰ):

[0006]

[0007] The novel polyphenolic substance 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone described in this invention was first isolated from kudzu root by the inventors.

[0008] The novel polyphenolic component 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone provided by this invention has been confirmed by in vitro pharmacological experiments to possess good alcohol dehydrogenase activating activity, with a half-maximal activation concentration (IC50) of 1.41 mM. The novel polyphenolic component 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone or its esterified derivatives can be used to prepare alcohol dehydrogenase activator drugs.

[0009] The novel polyphenolic component 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone or its esterified derivatives can be combined with excipients or carriers permitted in formulations or pharmaceuticals to prepare drugs or pharmaceutical compositions that can be used for hangover relief and liver protection. These drugs or pharmaceutical compositions can be in dosage forms such as wettable powders, tablets, granules, capsules, oral liquids, pellets, injections, and aerosols; they can also be in controlled-release or sustained-release dosage forms or nano-formulations known in the modern pharmaceutical industry.

[0010] Therefore, the present invention also provides an alcohol dehydrogenase activator drug comprising the above-mentioned novel polyphenolic component 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl) ethyl ketone or its esterified derivative as an active ingredient.

[0011] The alcohol dehydrogenase activator drug also includes excipients or carriers permitted by the formulation or drug, such as wettable powder, tablets, granules, capsules, oral liquid, pellets, injections, aerosols and other dosage forms; it can also adopt controlled-release or sustained-release dosage forms or nano-formulations known in the modern pharmaceutical industry.

[0012] This invention also provides a method for preparing a novel polyphenolic component, 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone, which is obtained by isolating from kudzu, wherein the kudzu can be wild kudzu or kudzu root.

[0013] The preferred steps for preparing the novel polyphenolic component 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone of the present invention are as follows: The root of *Pueraria lobata* is pulverized and extracted with a 95% ethanol aqueous solution. The extract is separated and the residue is collected. The residue is then added to a 2M NaOH solution (1:20, w / v) and extracted under nitrogen for 1 hour. Subsequently, the pH is adjusted to 1 with 6M HCl, and the extract is obtained by 5 extractions with ethyl acetate (1:1, v / v). The ethyl acetate phase is collected, concentrated under reduced pressure to remove the organic solvent, and the extract is obtained. The extract was subjected to normal-phase silica gel column chromatography, eluted with chloroform / methanol at a gradient ratio from 100:0 to 60:40 (v / v), and fraction F8 eluted at a chloroform / methanol volume ratio of 70:30 (v / v) was collected. Fraction F8 was subjected to ODS-A medium-pressure column chromatography, eluted with methanol / water / alcohol at a gradient ratio from 20:80 to 90:10 (v / v), and fraction F8-2 eluted at a methanol / water volume ratio of 30:70 (v / v) was collected. Fraction F8-2 was purified by high performance liquid chromatography to obtain 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone.

[0014] This invention extracts and isolates an alcohol dehydrogenase activator from kudzu root. The preparation process is controllable, convenient, environmentally friendly, and has potential economic benefits. The resulting novel polyphenolic compound, 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)acetone, is stable and easily stored. Its alcohol dehydrogenase activating activity is excellent, making it a promising lead compound for developing novel hepatoprotective drugs for alcohol detoxification. Attached Figure Description

[0015] Figure 1 It is the compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone. 1 H NMR spectrum;

[0016] Figure 2 It is the compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone. 13 C NMR spectrum;

[0017] Figure 3 It is the compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone. 1 H -1 HCOSY diagram;

[0018] Figure 4 The HSQC spectrum of compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone is shown.

[0019] Figure 5The HMBC spectrum of compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone is shown.

[0020] Figure 6 This is the HR-EI-MS spectrum of compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone. Detailed Implementation

[0021] The following embodiments are further illustrations of the present invention and are not intended to limit the present invention. Simple improvements made to the present invention based on the essence of the present invention are all within the scope of protection claimed by the present invention.

[0022] Example 1: Preparation of the polyphenol compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone

[0023] 1.1 Extraction and Separation

[0024] After slicing and crushing the kudzu root, it was extracted with 95% ethanol aqueous solution. The extract was separated and the residue was collected. The residue was then extracted with 2M NaOH solution (1:20, w / v g / ml) under nitrogen atmosphere for 1 hour. The pH was then adjusted to 1 with 6M HCl and extracted with ethyl acetate (1:1, v / v) 5 times. The ethyl acetate phase was collected and concentrated under reduced pressure to remove organic solvents, and the extract was obtained. The extract was subjected to normal-phase silica gel column chromatography, eluted with chloroform / methanol at a gradient ratio from 100:0 to 60:40 (v / v), and fraction F8 eluted at a chloroform / methanol volume ratio of 70:30 (v / v). Fraction F8 was then subjected to ODS-A medium-pressure column chromatography, eluted with methanol / water / alcohol at a gradient ratio from 20:80 to 90:10 (v / v), and fraction F8-2 eluted at a methanol / water volume ratio of 30:70 (v / v). Fraction F8-2 was further purified by preparative HPLC using a YMC-Pack ODS-A column (5 μm, 20 × 250 mm) to obtain compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)acetone (40 mg, tR 92 min).

[0025] 1.2 Structural Identification of New Polyphenolic Compounds

[0026] Figure 1 It is the compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone. 1 H NMR spectrum; Figure 2 It is 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone 13 C NMR spectrum; Figure 3It is the compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone. 1 H- 1 H COSY spectrum; Figure 4 The HSQC spectrum of compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone is shown. Figure 5 The HMBC spectrum of compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone is shown. Figure 6 This is the HR-EI-MS spectrum of compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone;

[0027] Table 1. NMR data of compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone (deuterated methanol assay)

[0028]

[0029] Based on the comprehensive analysis of the above mass spectrometry and one-dimensional and two-dimensional NMR spectroscopic data, the structural formula of the compound is derived as shown in formula (Ⅰ), and it is named 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone:

[0030]

[0031] Example 2: Detection of alcohol dehydrogenase activation activity of the polyphenol compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone

[0032] 0.1 mL of test samples of different concentrations were added to cuvettes containing 1.4 mL distilled water, 0.75 mL 1.0 M Tris-HCl buffer (pH 8.8), 0.3 mL 20 mm NAD+ solution, and 0.3 mL 0.2 M ethanol. A blank control group was also included (the sample solution was replaced with distilled water). After thorough mixing, the solution was incubated at 25 °C for 5 minutes. Immediately afterwards, 0.15 mL of ADH (5 U / mL) was added to the cuvette, and the absorbance at 340 nm was measured every 30 seconds using a Varian 100 UV spectrophotometer to calculate the increase in NADH absorption. The change in absorbance (A340 / min) was estimated based on a standard curve. The activation rate of ADH (U / L) was calculated as follows:

[0033] Activation rate (%) = (A1 - A0) / A0 × 100

[0034] A1 is the absorbance value of the sample group, and A0 is the absorbance value of the blank group.

[0035] The experimental data are shown in Table 2:

[0036] Table 2. Ethanol dehydrogenase activation activity of compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone

[0037]

[0038] 3. Experimental conclusions:

[0039] This experiment shows that the polyphenol compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone has good alcohol dehydrogenase activation activity, and it is expected to be developed for the preparation of hangover relief and liver protection drugs. It has broad application potential and promising prospects.

[0040] The above are merely preferred embodiments of the present invention. It should be noted that the above preferred embodiments should not be considered as limitations on the present invention, and the scope of protection of the present invention should be determined by the scope defined in the claims. For those skilled in the art, several improvements and modifications can be made without departing from the spirit and scope of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone or its esterified derivatives, the structure of which is shown in formula (Ⅰ):

2. A method for preparing the alkaloid compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone according to claim 1, characterized in that, The 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone was isolated from kudzu root.

3. The preparation method according to claim 2, characterized in that, The specific steps are as follows: a. Preparation of extract: The whole kudzu root powder was extracted with 95% ethanol aqueous solution, the extract was separated and the residue was collected. The residue was added to 2M NaOH solution and extracted under nitrogen atmosphere. Then the pH was adjusted to 1 with 6M HCl and extracted with ethyl acetate. The ethyl acetate phase was collected and concentrated under reduced pressure to remove organic solvents to obtain the extract. b. Separation and purification: The extract was subjected to normal-phase silica gel column chromatography, eluted with chloroform / methanol at a volume ratio of 100:0 to 60:40, and fraction F8 eluted at a volume ratio of 70:30 was collected. Fraction F8 was subjected to ODS-A medium-pressure column chromatography, eluted with methanol / water / alcohol at a volume ratio of 20:80 to 90:10, and fraction F8-2 eluted at a volume ratio of 30:70 was collected. Fraction F8-2 was purified by high performance liquid chromatography to obtain 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone.

4. The preparation method according to claim 3, characterized in that, The kudzu root mentioned is either kudzu root or wild kudzu, including the tuberous root, leaves, and stems of kudzu.

5. The use of the alkaloid compound 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl) ethyl ketone or its esterified derivative as described in claim 1 in the preparation of an alcohol dehydrogenase activator drug.

6. An alcohol dehydrogenase activator drug, characterized in that, Contains an effective amount of 1-(2-hydroxy,4-O-glucosylphenyl)-2-(4-hydroxyphenyl)ethyl ketone or its esterified derivative as an active ingredient, as described in claim 1.

7. The alcohol dehydrogenase activator drug according to claim 7, characterized in that, It also includes excipients or carriers permitted in the formulation or drug.

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