CN114380680B

CN114380680B - Flavonoid compound and application thereof

Info

Publication number: CN114380680B
Application number: CN202210065169.XA
Authority: CN
Inventors: 钱朝南; 李长志; 陈金东; 周红娟; 黄婕
Original assignee: Guangzhou Chaoliliang Biological Technology Co ltd
Current assignee: Guangzhou Chaoliliang Biological Technology Co ltd
Priority date: 2022-01-20
Filing date: 2022-01-20
Publication date: 2022-09-16
Anticipated expiration: 2042-01-20
Also published as: CN114380680A

Abstract

The invention belongs to the field of medicinal chemistry, and particularly relates to a flavonoid compound and application thereof. The flavonoid compound provided by the invention has a structure shown in a general formula I:

Figure RE-RE-RE-462048DEST_PATH_IMAGE001

(I) wherein, R1 is independently selected from H, OH, CH3, OCH3, Cl, NO 2; r2, R3 are independently selected from H, OH, CH 3; r4, R5 are independently selected from H and OH. The research shows that the compound with the structure of the general formula I has remarkable cell migration promoting capability, animal experiments further verify that the compound can remarkably promote wound healing, and the compound can be used for preparing medicines for promoting wound healing, particularly medicines for accelerating wound healing of diabetic patients.

Description

Flavonoid compound and application thereof

Technical Field

The invention belongs to the field of medicinal chemistry, and particularly relates to a flavonoid compound and application thereof.

Background

Tissue damage and repair are the hot subjects of current research in the surgical field, and clinically, repair disorders are mainly manifested by healing delayed skin wounds, such as diabetic ulcers, leg ulcers, pressure ulcers and the like, and are one of the difficult problems of clinical treatment due to high morbidity and high disability rate. The common clinical refractory disease of delayed wound healing always has difficulty in the medical field, and how to effectively promote wound healing is a hot problem to be solved urgently in the current medical science.

Cell motility is critical in embryonic development, tumor metastasis, wound healing and immune response, and has been reported: living cells will respond to biochemical signals, moving to high concentrations, and cells also present responding mechanical forces, and cells will react to chemical and mechanical signals using the same molecular network, combining potentially conflicting signals to form a uniform pathway. Some traditional Chinese medicines have good curative effect on chronic wounds, and except oral traditional Chinese medicines, the external traditional Chinese medicines play an important role in different stages of wound healing. However, the research on external traditional Chinese medicines is still in the beginning stage, and the basic research on the substances from the drug effect to the traditional Chinese medicines is being preliminarily explored. At present, the literature reports of external traditional Chinese medicines for promoting cell migration to promote wound healing are few. The existing medicine for promoting wound healing has the risk of tumor diffusion while promoting wound healing, and particularly for patients with tumor complicated with diabetes, so that the medicine capable of accelerating wound healing of patients without causing tumor cell diffusion is urgently needed.

Selene de Jes U.S. Acosta-Cota et al, in "Therapeutic effect of 4-hydroxychalcone in male wistatic fatty liver disease" (European Journal of Pharmacology,2019,863(C):172699.) disclose the Therapeutic effect of 4-hydroxychalcone on nonalcoholic fatty liver disease male Wistar rats.

Alshangici Amnah M, Tuboyy Ezter, Hegarty shade V, McCarthy Cathal M, Sullivan Aideen M, O' Keeffe Gerard W et al, 4-Hydroxychalcone Cell Death vision in MYCN-Amplified Human neuroplastoma Cells (Oxidative medium and cellular diversity, 2019,2019:) disclose that 4-Hydroxychalcone treatment significantly reduces intracellular glutathione antioxidant levels and increases intracellular reactive oxygen species. Meanwhile, when used in combination with anticancer drugs cisplatin or adriamycin, 4-hydroxychalcone induces a greater reduction in cell viability than when two anticancer drugs are used alone. However, it is not disclosed that the 4-hydroxychalcone compound has the effects of promoting cell migration and promoting wound healing.

Disclosure of Invention

The invention aims to provide a flavonoid compound and application thereof, and the flavonoid compound can be used for preparing a medicament for promoting wound healing, in particular to a medicament for accelerating wound healing of a diabetic patient.

In order to achieve the purpose, the invention adopts the following technical scheme: a flavonoid compound having the structure of formula I:

wherein R is ₁ Independently selected from H, OH, CH ₃ 、OCH ₃ 、Cl、NO ₂ ；

R ₂ 、R ₃ Independently selected from H, OH, CH ₃ ；

R ₄ 、R ₅ Independently selected from H and OH.

Preferably, the flavonoid comprises the structure:

preferably, the compound having the structure of formula I is 4-hydroxychalcone, having the structure:

the invention also provides application of the flavonoid compound in preparing a medicament for promoting wound healing.

Preferably, the medicament further comprises one or more pharmaceutically acceptable carriers or pharmaceutic adjuvants.

Preferably, the medicament is a liquid, a paste, a gel or a dressing.

The invention also provides application of the flavonoid compound in preparing a medicament for promoting movement of fibroblasts.

The invention also provides application of the flavonoid compound in preparing a medicament for promoting wound healing of a diabetic surgical patient.

Compared with the prior art, the invention has the following beneficial effects:

(1) according to the invention, cell scratch experiments show that the flavonoid compound with the general formula (I) has obvious cell migration promoting capability under a safe dosage, wherein the 4-hydroxychalcone has the best effect.

(2) According to the invention, through wound healing experiments of normal mice and diabetic mice, the flavonoid compounds with the general formula (I) are found to have obvious healing effects on skin wounds of the mice for the first time, especially have obvious treatment effects on diabetic patients with wounds which are difficult to heal, and the 4-hydroxychalcone has the best wound healing effect and can be used for preparing medicines for promoting wound healing.

Drawings

FIG. 1 is a schematic diagram showing the effect of 4-hydroxychalcone of the present invention in promoting the movement scratch test of 3T 3.

FIG. 2 is a graph showing data of a 4-hydroxychalcone-promoted 3T3 sports scratch test of the compound of the present invention.

FIG. 3 is a schematic diagram showing the healing effect of 4-hydroxychalcone of the present invention on wounds of normal mice.

FIG. 4 is a graph showing the data on the healing rate of 4-hydroxychalcone of the present invention on the wound of a normal mouse at day 9.

FIG. 5 is a schematic diagram showing the healing effect of 4-hydroxychalcone of the compound of the present invention on wounds of diabetic mice.

FIG. 6 is a graph showing the wound healing rate data of diabetic mice on day 11 of the compound 4-hydroxychalcone of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1 cell scratch test

1.1 subjects: mouse embryonic fibroblast 3T 3.

1.2 Experimental drugs: 4-hydroxy chalcone and a negative control reagent 0.1% DMSO.

1.3 Experimental methods: and (3) uniformly scribing by using a marker thin head behind the 6-hole plate and using a ruler, traversing the through holes approximately every 0.5-1cm, generally scribing three lines, and sequentially naming the lines as a, b and c. The line b crosses the midpoint, and the other two lines are equally spaced on both sides. Pre-plating 3T3 cells were pre-treated with 4-hydroxychalcone (5. mu.M) for about 24 h. In six-well plate, each well is 2ml, and each well is added with 1.5X 10 ⁶ 3T3 cells, each of which was plated with 2 wells, were cultured in an incubator. The cell number is preferably more than 90% of the cells can be spread on the wall overnight, and the cell number is properly adjusted. Opening the cover of the orifice plate, sucking off the old culture medium, putting a ruler vertical to the b-line frame on the orifice plate, using 200ul of gun head to cling to the ruler to move uniformly to manufacture a cell scribing line, likewise, scribing two parallel lines at equal intervals on two sides of the line, and respectively naming the lines as line 1, line 2 and line 3 from left to right. Gently washing the cells with sterile 1 × PBS for 3 times, removing the scraped cells, adding serum culture medium, adding 4-hydroxychalcone (working concentration of 10 μ M), and adding 5% CO at 37 deg.C ₂ An incubator. The samples were taken and photographed at the time of dosing 0, 24h, 48 h.

1.4 results of the experiment

The experimental results are shown in fig. 1 and fig. 2, and compared with the negative control group, the 4-hydroxychalcone compound has a remarkable promoting effect on the migration of the mouse embryonic fibroblast 3T3 no matter the compound acts for 24 hours or 48 hours.

Example 2 Effect on healing of wounds in mice

2.1 Experimental group design

TABLE 1 experimental group design

2.2 construction of diabetic mouse model with high fat diet in combination with STZ

(1) Mouse ordering: ordering male BALB/c mice of 3 weeks old from the medical experimental animal center of Guangdong province, feeding in an SPF feeding room for 5 days, and performing isolation observation;

(2) the first day of the experiment was the start of high fat diet feeding of the mice in the model group. Recording body weight weekly, observing hair, food intake and urination, removing feed at night 28 days, and fasting overnight;

(3) preparing Streptozotocin (STZ) injection: 20mg of STZ powder was dissolved in 4mL of citric acid buffer solution to prepare an injection solution of 5 mg/mL. All operations are carried out on ice, attention is paid to light protection, and the STZ solution is prepared for use immediately;

(4) injection of STZ: the following day, mice body weight and fasting glucose were measured: injecting STZ solution (40mg/kg/d) into abdominal cavity of model mouse once a day for 5 days continuously, continuously fasting the first day, and resuming high fat feeding the second day; continuously observing the feeding, drinking and urination conditions of the mice;

(5) after 2 weeks from the end of dosing, two groups of mice were tested for fasting blood glucose:

1) in the modeling mice, the mice which are stable for more than 3 times at more than 11.1mmoL/L and appear 'polydipsia, polyuria, polyphagia and emaciation' are defined as the diabetes mice which are successfully modeled, the mice are included in the subsequent experiments, the mice which do not meet the standard of the model exit the experiments, and the modeling success rate is counted;

2) in the non-model group mice, the stability is not more than 11.1mmoL/L for more than 3 times continuously, no mice with the expression of polydipsia, diuresis, polyphagia and emaciation are brought into the subsequent experiment, and the mice which do not meet the model standard quit the experiment;

(6) during the experiment, the sanitary nursing of the model-making mice is noticed, and if polydipsia and polyuria appear, the drinking water must be supplemented in time, so that the interval for replacing padding is shortened.

2.3 mouse dorsal skin full-thickness wound construction and Observation of effects of Compounds on skin wound healing

a. Grouping: randomly dividing the diabetes group successfully modeled into a negative control group and a dosing group; the mice in the normal blood sugar group are randomly divided into a solvent control group and an administration group;

b. after successful modeling for 2 weeks, 1% sodium pentobarbital was anesthetized by intraperitoneal injection (7 ul/g);

c. placing the fully anesthetized mouse on a pad towel in a prone position, and removing back hair by using depilatory cream;

d. disinfecting back skin with 75% alcohol, respectively making 2 round wounds with diameter of about 15mm on two sides of the highest position of the midline of the back by using trephines with diameter of 15mm, removing subcutaneous tissue with surgical scissors and forceps, and exposing fascia on the surface of muscle;

e. stopping bleeding and disinfecting the wound, and opening the wound; taking a ruler as a control, and taking a picture to record the shape and size of the wound;

f. administration: each mouse in the control group was injected with the same volume (0.1 ml) of normal saline containing 10% DMSO-40% PEG 400-5% tween 80-45%; normal mice were administered 4-hydroxychalcone (30mg kg) ^-1 ·d ^-1 ) The diabetic mice were administered 4-hydroxychalcone (10 mg. kg) ^-1 ·d ^-1 ). The administration mode is intraperitoneal injection. The day of the mouse trauma is D1, the mice in each group are injected with D1, D3, D5, 7, 9, 11 and 13 respectively, and the wounds are photographed at D1, D3, D5, D7, D9, D11 and D13 to record the sizes;

g. the anesthetized mice were resuscitated under the temperature control plate and returned to the EVC cage of the clean rearing room.

2.4 results of the experiment

As shown in FIGS. 3, 4, 5 and 6, the results of the experiment were obtained by administering 30 mg/kg of 4-hydroxychalcone ^-1 ·d ^-1 Compared with a solvent control group, the wound healing rate of a normal mouse is remarkably improved within 3-9 days after administration, the wound healing rate of the mouse reaches 80% at the 9 th day, meanwhile, the wound healing rate of a diabetic mouse treated by the 4-hydroxychalcone compound is remarkably improved within 3-9 days after administration, and the situation that the wound is basically healed already appears at the 13 th day, so that the 4-hydroxychalcone compound has the effect of remarkably promoting the wound healing of the mouse.

The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.

Claims

Use of 4-hydroxychalcone for the preparation of a medicament for promoting fibroblast motility, characterized in that the structure of said 4-hydroxychalcone is as follows:

。
use of 4-hydroxy chalcone for the preparation of a medicament for promoting wound healing in a diabetic patient, wherein the 4-hydroxy chalcone has the following structure:

。
3. the use according to claim 1 or 2, wherein the medicament further comprises one or more pharmaceutically acceptable carriers or pharmaceutical excipients.
4. Use according to claim 3, wherein the medicament is a liquid, a paste, a gel or a dressing.