Application of quercetin-3-O-glucuronide in the preparation of drugs for promoting angiogenesis

Abstract

The present invention discloses the use of quercetin-3-O-glucuronide in the preparation of a drug that promotes angiogenesis, belonging to the technical field of Chinese herbal medicine development. The present invention utilizes the polarity characteristics of quercetin-3-O-glucuronide to isolate and obtain quercetin-3-O-glucuronide from Cyclocarya paliurus. Experiments in a zebrafish vascular injury model demonstrate that the quercetin-3-O-glucuronide provided by the present invention has good pro-angiogenic activity, showing potential for the preparation of drugs or functional foods that promote angiogenesis, resist damage, and prevent complications associated with diabetic vascular injury.

Description

Technical Field

[0001] The invention belongs to the technical field of development of Chinese herbal medicines, and particularly relates to the application of quercetin-3-O-glucuronide in the preparation of a drug for promoting angiogenesis. Background Art

[0002] Cyclocarya paliurus Cyclocarya paliurus (Batal) Iljinsk Cyclocarya paliurus, a tall tree belonging to the Juglandaceae family, is widely distributed in China's subtropical regions. It is a plant with both medicinal and edible properties, earning it the title of "the third tree of medicine." Cyclocarya paliurus tea, made from its buds and leaves, is effective in treating the "three highs," or high blood sugar, high blood sugar, and high cholesterol. Flavonoids are the primary indicative components of Cyclocarya paliurus. Cyclocarya paliurus flavonoids exhibit pharmacological effects such as hypoglycemic, antioxidant, antibacterial, anti-acute liver damage, and intestinal microbial regulation. The "Compendium of Chinese Materia Medica Resources" states that both the leaves and bark of Cyclocarya paliurus are medicinal, offering analgesic, heat-clearing, and detoxifying properties, and are used to treat stubborn ringworms. A polysaccharide (CP) extracted from Cyclocarya paliurus leaves was modified to yield its acetyl derivative, Ac-CP. Ac-CP significantly stimulated bacteriophage proliferation, exhibiting a significantly stronger effect than the corresponding unmodified CP. The polysaccharide from Cyclocarya paliurus significantly inhibited the viability of hypoxic A549 and H520 cells.

[0003] The natural ingredient quercetin-3-O-glucuronide ( Miquelianin ) compound is a CBR1 inhibitor. In our study, we found that the natural ingredient quercetin-3-O-glucuronide ( Miquelianin ) compounds have pharmacological activity in promoting angiogenesis. To date, there are no patents or literature reports on the research results of quercetin-3-O-glucuronide in promoting angiogenesis. Summary of the Invention

[0004] In order to overcome the shortcomings and deficiencies of the prior art, the present invention provides the use of quercetin-3-O-glucuronide in the preparation of a drug for promoting angiogenesis.

[0005] Quercetin-3-O-glucuronide is a flavonoid chemical and a natural component found in many plants. Bupleurum scorzonerifolium Willd. 、Bar Plate Return Polygonum perfoliatum L. , Cyclocarya paliurus Cyclocarya paliurus (Batalin) Iljinsk. In plants such as.

[0006] In order to achieve the above object, the present invention adopts the following technical means:

[0007] A first aspect of the present invention provides use of quercetin-3-O-glucuronide in the preparation of a drug for promoting angiogenesis. The drug comprises quercetin-3-O-glucuronide and a pharmaceutically acceptable carrier.

[0008] The second aspect of the present invention provides the use of quercetin-3-O-glucuronide in the preparation of a medicament for preventing complications associated with diabetic vascular damage. The medicament comprises quercetin-3-O-glucuronide and a pharmaceutically acceptable carrier.

[0009] The third aspect of the present invention provides the use of a pharmaceutical composition comprising quercetin-3-O-glucuronide as an active ingredient in the preparation of a drug for promoting angiogenesis.

[0010] A fourth aspect of the present invention provides the use of a pharmaceutical composition comprising quercetin-3-O-glucuronide as an active ingredient in the preparation of a medicament for preventing complications associated with diabetic vascular damage.

[0011] In some embodiments of the present invention, the types of pharmaceutical preparations include injections, drip solutions, powder injections, granules, tablets, granules, powders, oral solutions, sugar-coated tablets, film-coated tablets, enteric-coated tablets, buccal preparations, granules, pills, ointments, pills, sprays, dropping pills, disintegrants, orally disintegrating tablets, and pellets.

[0012] As used herein, a "pharmaceutically acceptable" ingredient is one that is suitable for use in humans and / or mammals without excessive adverse side effects (e.g., toxicity), i.e., a substance with a reasonable benefit / risk ratio. The term "pharmaceutically acceptable carrier" refers to a carrier used to administer a therapeutic agent, including various excipients and diluents. The term refers to pharmaceutical carriers that are not themselves essential active ingredients and are not unduly toxic upon administration. Suitable carriers are well known to those of ordinary skill in the art. A comprehensive description of pharmaceutically acceptable carriers can be found in Remington's Pharmaceutical Sciences (Mack Pub. Co., N.J. 1991).

[0013] Pharmaceutically acceptable carriers may contain liquids such as water, saline, glycerol, and sorbitol in pharmaceutical preparations. Additionally, auxiliary substances such as lubricants, glidants, wetting agents or emulsifiers, pH buffers, and stabilizers may also be present in these carriers.

[0014] A seventh aspect of the present invention provides the use of quercetin-3-O-glucuronide in the preparation of skin repair skin care products and / or cosmetics; for preventing subcutaneous blood vessel damage and promoting angiogenesis during skin repair.

[0015] Beneficial effects of the present invention

[0016] Compared with the prior art, the present invention has the following beneficial effects: the present invention provides quercetin-3-O-glucuronide (Miquelianin) which has pro-angiogenic pharmacological activity and can be used to prepare pro-angiogenic and anti-injury drugs, skin care products or cosmetics. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 is the chemical structure of quercetin-3-O-glucuronide;

[0018] Figure 2 is the H NMR spectrum of quercetin-3-O-glucuronide;

[0019] Figure 3 is the C NMR spectrum of quercetin-3-O-glucuronide;

[0020] Figure 4 is the IR spectrum of quercetin-3-O-glucuronide;

[0021] Figure 5 This is the comparison result of the vascular ISV length in different groups in the quercetin-3-O-glucuronide angiogenic activity experiment. DETAILED DESCRIPTION

[0022] The following examples are provided to illustrate preferred embodiments of the present invention. Those skilled in the art will appreciate that the techniques disclosed in the following examples represent techniques discovered by the inventors that can be used to practice the present invention and, therefore, can be considered preferred embodiments of the present invention. However, those skilled in the art will appreciate from this disclosure that many modifications may be made to the specific embodiments disclosed herein while still achieving the same or similar results without departing from the spirit or scope of the present invention.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which this invention belongs, and the disclosures herein and the materials they cite are hereby incorporated by reference. Those skilled in the art will recognize or be able to ascertain, through routine experimentation, many technical equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the claims.

[0024] The technical solution of the present application will be further described in detail below in conjunction with specific implementation methods.

[0025] Example 1 Preparation of Quercetin-3-O-glucuronide

[0026] Ten kg of leaves, rhizomes, and bark of Cyclocarya paliurus (Batal) Iljinsk were extracted with 10 L of methanol for two weeks. The methanol extract was concentrated and suspended in 1 L of distilled water. The aqueous suspension was extracted three times with 1 L of ethyl acetate. The ethyl acetate extract was concentrated to obtain 23 g of an extract. The sample was mixed with silica gel (100 mesh, 100 g) and subjected to normal phase silica gel column chromatography (200-300 mesh, 1 kg; silica gel column size L 500 mm, Ø 120 mm). Gradient elution was performed with a petroleum ether / ethyl acetate mixture of volume ratios of 9:1, 4:1, 3:1, 1:1, 1:3, and 1:9, respectively, with each gradient elution taking 5 L. Fractions were analyzed by TLC. Fractions with a 4:1 ratio were pooled, concentrated, and recrystallized from acetone (room temperature) to obtain the target compound, quercetin-3-O-glucuronide.

[0027] Example 2 Structural Identification of Quercetin-3-O-Glucuronide

[0028] The target compound prepared in Example 1 was purified by HPLC. The structure of the sample with a purity greater than 98% was determined by mass spectrometry and nuclear magnetic resonance. The nuclear magnetic resonance was determined by a Bruker AVANCE DRX-500 NMRSectrometer with TMS as the internal standard; the high-resolution mass spectrometry FTICRMS was determined by a Bruker Apex Spectrometer; and the electrospray ionization mass spectrometry ESI-MS was determined by a Bruker Esquire 3000 plus Spectrometer determination.

[0029] The chemical structure of the compound quercetin-3-O-glucuronide is as follows Figure 1 As shown, the H NMR spectrum of the compound quercetin-3-O-glucuronide is as follows Figure 2 As shown, the NMR C spectrum of the compound quercetin-3-O-glucuronide is as follows Figure 3 As shown, the IR spectrum of the compound quercetin-3-O-glucuronide is as follows Figure 4 The NMR data are shown in Table 1 below.

[0030] Table 1 NMR data of quercetin-3-O-glucuronide

[0031]

[0032] Example 3 Analysis of the angiogenic activity of quercetin-3-O-glucuronide

[0033] PTK787 (a specific vascular endothelial growth factor (VEGF) receptor kinase inhibitor), ginsenoside Rg1, and the compound were dissolved in distilled water containing 1% DMSO. After 48 hours, the selected fluorescent zebrafish eggs were placed in a 96-well plate to set up the experimental groups, namely:

[0034] (1) Blank control group: E3+PTU culture medium supplemented with 1% DMSO;

[0035] (2) Model injury group and natural repair group: 0.25 μg / mL PTK787 was used to create model injury;

[0036] (3) Positive control group: 0.25 μg / mL PTK787 was used to create injury models;

[0037] (4) Experimental drug group: 0.25 μg / mL PTK787 was used to create injury models;

[0038] After 72 hours, the damage of zebrafish in the above four groups was observed under a fluorescence microscope and the following operations were performed respectively;

[0039] (2) Model injury group and natural repair group: only add the same amount of E3 culture medium and perform natural repair;

[0040] (3) Positive control group: 100 μg / mL ginsenoside Rg1 was added for repair;

[0041] (4) Experimental drug group: 200 μL of 40 μg / mL experimental drug: quercetin-3-O-glucuronide was added for repair;

[0042] After 96 hours, each zebrafish was photographed under a fluorescence microscope using methylcellulose as a fixative. The above operation was repeated 6 times to determine the angiogenic activity of these compounds. Zebrafish vascular images were processed using Image Pro Plus 5.0 and the length of angiogenesis was measured. The results of the 6-repetition operation are shown in Table 2; the average values ​​of the four groups are compared as shown in Table 2. Figure 5 shown.

[0043] Table 2 Analysis of angiogenic activity of quercetin-3-O-glucuronide in zebrafish model

[0044]

[0045] The results showed that the positive control group, which received ginsenoside Rg1 for repair, saw a 9.3% increase in ISV length compared to the naturally repaired model injury group. The experimental group, which received the compound quercetin-3-O-glucuronide for repair, saw an 18.9% increase in ISV length compared to the naturally repaired model injury group. Compared to the blank control group, the effect of ginsenoside Rg1 repair was inferior to the natural growth in the blank control. The experimental group, which received the compound quercetin-3-O-glucuronide for repair, saw a 1.5% increase in ISV length compared to the natural growth in the blank control. These results suggest that quercetin-3-O-glucuronide (Miquelianin) has pro-angiogenic pharmacological activity.

[0046] Example 4 Preparation of Quercetin-3-O-Glucuronide-Containing Dropping Pills

[0047] Take 0.5 g of quercetin-3-O-glucuronide and polyethylene glycol-6000, mix them evenly, heat to melt, and transfer the materials to the drip irrigation system. The drug solution is dripped into 6-8℃ liquid paraffin, and the oil is removed to prepare 300 dripping pills.

[0048] Example 5 Preparation of quercetin-3-O-glucuronide freeze-dried powder injection

[0049] Take 0.5 g of quercetin-3-O-glucuronide, 4.5 g of glucose, 0.9 g of sodium thiosulfate and 1000 mL of distilled water, mix the above components evenly, freeze-dry, and package into 400 vials to obtain quercetin-3-O-glucuronide freeze-dried powder injection.

[0050] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the present application.

Claims

1. Use of quercetin-3-O-glucuronide in the preparation of a medicament for preventing diabetic vascular damage and related complications, characterized in that: The drug comprises quercetin-3-O-glucuronide and pharmaceutically acceptable excipients, and can promote angiogenesis.

2. Use of a pharmaceutical composition containing quercetin-3-O-glucuronide as an active ingredient in the preparation of a medicament for preventing diabetic vascular damage and related complications, characterized in that: The drug can promote angiogenesis.

3. The use according to any one of claims 1-2, characterized in that: The preparation types of the drug include injection, granules, tablets, pills, ointments, pills, and sprays.

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