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

Abstract

The invention discloses application of quercetin-3-O-glucuronide in preparation of a medicine for promoting angiogenesis, and belongs to the technical field of Chinese herbal medicine development. According to the application, the quercetin-3-O-glucuronide is separated from cyclocarya paliurus by utilizing the polarity characteristic of the quercetin-3-O-glucuronide, and a zebra fish vascular injury model test shows that the quercetin-3-O-glucuronide provided by the invention has relatively good angiogenesis promoting activity, and can be used for preparing a medicine for treating the angiogenesis. The invention has a development prospect in the aspect of preparing medicines or functional foods for promoting angiogenesis, resisting injury and preventing diabetic vascular injury related complications.

Description

Technical Field

[0001] The invention belongs to the technical field of development of Chinese herbal medicines, and specifically 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 , belongs to the genus Cyclocarya paliurus of the Juglandaceae family. It is a tall tree widely distributed in the subtropical regions of China. It is a plant with both medicinal and edible properties and is known as the "third tree in the medical field". The buds and leaves of Cyclocarya paliurus tea have a significant effect in treating the "three highs". Flavonoids are the main indicator components of Cyclocarya paliurus. Cyclocarya paliurus flavonoids have pharmacological effects such as hypoglycemic, antioxidant, antibacterial, anti-acute liver damage, and intestinal microbial regulation. It is recorded in the "Chinese Traditional Medicine Resources" that both the leaves and skin of Cyclocarya paliurus can be used as medicine, with the effects of analgesia, clearing away heat and detoxification, and are used to treat stubborn ringworms; the polysaccharide (CP) extracted from the leaves of Cyclocarya paliurus is modified to obtain its acetyl derivative Ac-CP. Ac-CP can significantly stimulate the proliferation of bacteriophages, and its effect is significantly stronger than that of the corresponding unmodified polysaccharide CP. The polysaccharides in Cyclocarya paliurus have significantly inhibited the viability of hypoxic A549 and H520 cells.

[0003] Quercetin-3-O-glucuronide (quercetin-3-O-glucuronide) is a natural ingredient extracted and isolated from Cyclocarya paliurus. Miquelianin ) compound is a CBR1 inhibitor. In our study, we found that the natural ingredient quercetin-3-O-glucuronide ( Miquelianin ) compounds have pharmacological activity to promote angiogenesis. So far, 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. 、Guard Plate Return Polygonum perfoliatum L. , Cyclocarya paliurus Cyclocarya paliurus (Batalin) Iljinsk. And other plants.

[0006] In order to achieve the above object, the present invention adopts the following technical means: The first aspect of the present invention provides the 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.

[0007] The second aspect of the present invention provides the use of quercetin-3-O-glucuronide in the preparation of a medicament for preventing complications related to diabetic vascular damage. The medicament comprises quercetin-3-O-glucuronide and a pharmaceutically acceptable carrier.

[0008] 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.

[0009] The 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.

[0010] 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, lozenges, granules, pills, ointments, pills, sprays, drop pills, disintegrants, orodisintegrating tablets, and pellets.

[0011] As used herein, "pharmaceutically acceptable" ingredients are suitable for use in humans and / or mammals without excessive adverse side effects (such as toxicity), i.e., substances with a reasonable benefit / risk ratio. The term "pharmaceutically acceptable carrier" refers to a carrier used for administering a therapeutic agent, including various excipients and diluents. The term refers to such pharmaceutical carriers: they are not necessary active ingredients themselves and are not excessively toxic after administration. Suitable carriers are well known to those of ordinary skill in the art. A full description of pharmaceutically acceptable carriers can be found in Remington's Pharmaceutical Sciences (Mack Pub. Co., N. J. 1991).

[0012] Pharmaceutically acceptable carriers may contain liquids such as water, saline, glycerol and sorbitol in pharmaceutical preparations. In addition, auxiliary substances such as lubricants, glidants, wetting agents or emulsifiers, pH buffer substances and stabilizers may also be present in these carriers.

[0013] The fifth aspect of the present invention provides the use of quercetin-3-O-glucuronide in the preparation of a health food for preventing vascular damage. The health food comprises quercetin-3-O-glucuronide and a food-acceptable auxiliary material.

[0014] The sixth aspect of the present invention provides the use of quercetin-3-O-glucuronide in the preparation of a health food for preventing complications related to diabetic vascular damage. The health food comprises quercetin-3-O-glucuronide and a food-acceptable auxiliary material.

[0015] The 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 vascular damage and promoting angiogenesis during skin repair.

[0016] Beneficial effects of the present invention 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 anti-injury drugs, preventive health foods, skin care products or cosmetics. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 is the chemical structure of quercetin-3-O-glucuronide; Figure 2 is the H NMR spectrum of quercetin-3-O-glucuronide; Figure 3 is the C NMR spectrum of quercetin-3-O-glucuronide; Figure 4 is the IR spectrum of quercetin-3-O-glucuronide; Figure 5 This is the comparison result of vascular ISV length in different groups in the quercetin-3-O-glucuronide angiogenic activity experiment. DETAILED DESCRIPTION

[0018] The following examples are used to demonstrate preferred embodiments of the present invention. It will be appreciated by those skilled in the art that the techniques disclosed in the following examples represent techniques discovered by the inventors that can be used to implement the present invention and therefore can be considered as preferred embodiments of the present invention. However, it will be appreciated by those skilled in the art based on this specification that many modifications may be made to the specific embodiments disclosed herein and still achieve the same or similar results without departing from the spirit or scope of the present invention.

[0019] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs, and the materials cited herein and those cited by them are incorporated by reference. Those skilled in the art will recognize or will learn through routine experimentation that there are many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the claims.

[0020] The technical solution of the present application is further described in detail below in conjunction with specific implementation methods.

[0021] Example 1 Preparation of Quercetin-3-O-glucuronide 10 kg of leaves, rhizomes and barks of Cyclocarya paliurus (Batal) Iljinsk were extracted with 10 L of methanol for 2 weeks. The methanol extract was concentrated and suspended with 1 L of distilled water. The water suspension was extracted three times with 1 L of ethyl acetate. The ethyl acetate extract was concentrated to obtain 23 g of 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 with a volume ratio of 9:1, 4:1, 3:1, 1:1, 1:3, and 1:9, respectively, with each gradient elution of 5 L. The fractions were detected by TLC, and the fractions with an elution ratio of 4:1 were collected, combined, concentrated, and then recrystallized with acetone (room temperature) to obtain the target compound quercetin-3-O-glucuronide.

[0022] Example 2 Structural Identification of Quercetin-3-O-Glucuronide The target compound prepared in Example 1 was identified for purity by HPLC. The sample with a purity greater than 98% was identified for structure by mass spectrometry and nuclear magnetic resonance techniques. The nuclear magnetic resonance was determined by Bruker AVANCE DRX-500 NMRSectrometer with TMS as internal standard; the high-resolution mass spectrometry FTICRMS was determined by Bruker Apex Spectrometer; and the electrospray mass spectrometry ESI-MS was determined by Bruker Esquire 3000 plus Spectrometer determination.

[0023] 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 C NMR 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.

[0024] Table 1 NMR data of compound quercetin-3-O-glucuronide

[0025] Example 3 Analysis of the angiogenic activity of quercetin-3-O-glucuronide PTK787 (specific VEGF receptor kinase inhibitor), ginsenoside Rg1 and compounds 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, which were: (1) Blank control group: E3+PTU culture medium supplemented with 1% DMSO; (2) Model injury group and natural repair group: 0.25 μg / mL PTK787 was used for model injury; (3) Positive control group: 0.25 μg / mL PTK787 was used to create injury models; (4) Experimental drug group: 0.25 μg / mL PTK787 was used to create injury models; After 72 h, the above four groups were observed for zebrafish damage under a fluorescence microscope and performed the following operations respectively; (2) Model injury group and natural repair group: only add an equal amount of E3 culture medium to allow natural repair; (3) Positive control group: Add 100 μg / mL ginsenoside Rg1 for repair; (4) Experimental drug group: Add 200 μL of 40 μg / mL experimental drug: quercetin-3-O-glucuronide for repair; After 96 hours, methylcellulose was used as a fixative, and each zebrafish was photographed under a fluorescence microscope. The above operation was repeated 6 times to determine and verify the angiogenic activity of these compounds. Image Pro Plus5.0 was used to process the zebrafish vascular images and measure the length of angiogenesis. The results of the 6-times repeated operation are shown in Table 2; the average comparison results of the four groups are shown in Figure 5 shown.

[0026] Table 2 Analysis of angiogenic activity of quercetin-3-O-glucuronide in zebrafish model

[0027] The results showed that the positive control group repaired with ginsenoside Rg1 increased the length of vascular ISV by 9.3% compared with the natural repair group of model injury, and the experimental drug group repaired with compound quercetin-3-O-glucuronide test drug increased the length of vascular ISV by 18.9% compared with the natural repair group of model injury. Compared with the blank control group, the effect of repairing with ginsenoside Rg1 was not as good as the natural growth in the blank control, and the experimental drug group repaired with compound quercetin-3-O-glucuronide test drug increased the length of vascular ISV by 1.5% compared with the natural growth in the blank control. The results showed that quercetin-3-O-glucuronide (Miquelianin) has pharmacological activity to promote angiogenesis.

[0028] Example 4 Preparation of Quercetin-3-O-glucuronide Dropping Pills 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 of pills. The drug solution is dripped into 6-8℃ liquid paraffin, degreased, and 300 pills are prepared.

[0029] Example 5 Preparation of quercetin-3-O-glucuronide freeze-dried powder injection 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 them, and pack them into 400 vials to obtain quercetin-3-O-glucuronide freeze-dried powder injection.

[0030] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference separately. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined in the present application.

Claims

1. Use of quercetin-3-O-glucuronide in the preparation of a drug for promoting angiogenesis, characterized in that: The medicine comprises quercetin-3-O-glucuronide and pharmaceutically acceptable excipients.

2. Use of quercetin-3-O-glucuronide in the preparation of a drug for preventing complications related to diabetic vascular damage, characterized in that: The medicine comprises quercetin-3-O-glucuronide and pharmaceutically acceptable excipients.

3. Use of a pharmaceutical composition comprising quercetin-3-O-glucuronide as an active ingredient in the preparation of a drug for promoting angiogenesis.

4. Use of a pharmaceutical composition comprising quercetin-3-O-glucuronide as an active ingredient in the preparation of a drug for preventing complications associated with diabetic vascular damage.

5. The use according to any one of claims 1 to 4, characterized in that: The preparation types of the drug include injection, drip, powder injection, granules, tablets, granules, powders, oral liquid, sugar-coated tablets, film-coated tablets, enteric-coated tablets, lozenges, granules, pills, ointments, pills, sprays, dropping pills, disintegrants, orodisintegrating tablets, and micropills.

6. Use of quercetin-3-O-glucuronide in the preparation of a health food for preventing vascular damage, characterized in that: The health food comprises quercetin-3-O-glucuronide and auxiliary materials acceptable to food.

7. Use of quercetin-3-O-glucuronide in the preparation of a health food for preventing complications related to diabetic vascular damage, characterized in that: The health food comprises quercetin-3-O-glucuronide and auxiliary materials acceptable to food.

8. Use of quercetin-3-O-glucuronide in the preparation of skin repairing skin care products and / or cosmetics.

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