CN113768941A

CN113768941A - Application of 2-deoxy-D-glucose in preparation of medicine for treating chronic eczema

Info

Publication number: CN113768941A
Application number: CN202111188077.2A
Authority: CN
Inventors: 张泉龙; 秦路平; 程真真; 张巧艳
Original assignee: Zhejiang Chinese Medicine University ZCMU
Current assignee: Zhejiang Chinese Medicine University ZCMU
Priority date: 2021-10-12
Filing date: 2021-10-12
Publication date: 2021-12-10

Abstract

The invention discloses an application of 2-deoxy-D-glucose in preparing a medicament for treating chronic eczema, wherein the 2-deoxy-D-glucose can be independently administered or added with pharmaceutic adjuvant in treating chronic eczema, and the 2-deoxy-D-glucose and other medicaments are combined and added with adjuvant to prepare various preparation formulations; the prepared preparation can be external preparation such as ointment, gel and the like, and can also be oral preparation.

Description

Application of 2-deoxy-D-glucose in preparation of medicine for treating chronic eczema

Technical Field

The invention relates to application of 2-deoxy-D-glucose serving as a medicament, in particular to application of 2-deoxy-D-glucose in preparation of a medicament for treating chronic eczema.

Background

Eczema (Eczema) is an inflammation reaction of skin with obvious exudation tendency caused by various factors, and is characterized in that the rash forms are various, and the rash in a chronic stage is usually characterized by local infiltration and hypertrophy, pruritus and repeated attack, and is a common skin disease in clinic. Factors that play a role in the pathogenesis of eczema are: interactions between genetics and environment, skin barrier dysfunction, microbial imbalances, immune disorders, and environmentally induced skin inflammation. It is presently believed that the direct mechanism of eczematous lesions is inflammation associated with Th1/Th2 cell dysregulation. Currently, topical or systemic medications are mainly used with glucocorticoids, traditional Chinese medicines, antibacterial drugs, and the like. At present, glucocorticoid can inhibit inflammatory reaction, is a common medicament for treating dermatitis and eczema, but has obvious adverse reaction in local or systemic administration, and the phenomenon of skin damage aggravation after medicament withdrawal can occur in clinical treatment, so the clinical application is greatly limited.

2-deoxy-D-glucose (2-deoxy-D-glucose) is a glucose analog, an inhibitor of glucose metabolism, which acts on hexokinase (hexokinase) to inhibit glycolysis. Has the effects of interfering the synthesis of virus specific glycoprotein, effectively inhibiting herpes simplex virus, RNA and DNA enveloped virus, breast cancer cell proliferation and the like. Antiviral and anticancer drugs prepared from 2-deoxy-D-glucose have been used in clinical treatment studies of herpes simplex virus, influenza, cancer and the like, and 2-deoxy-D-glucose also has an anti-rheumatoid arthritis effect, but reports of 2-deoxy-D-glucose for treating chronic eczema are not available.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides application of 2-deoxy-D-glucose in preparing a medicament for treating chronic eczema.

The purpose of the invention is realized by the following technical scheme: a new use of 2-deoxy-D-glucose in preparing medicine for treating chronic eczema is disclosed, which contains 2-deoxy-D-glucose and medicinal carrier.

The invention has the beneficial effects that: the 2-deoxy-D-glucose has the functions of regulating the balance of chronic eczema Th1/Th2 cells, inhibiting inflammatory reaction and improving the ear swelling of mice.

Drawings

FIG. 1 is a chemical structural diagram of 2-deoxy-D-glucose;

FIG. 2 is a graph showing the effect of 2-deoxy-D-glucose on the difference in weight of ears of mice with chronic eczema;

FIG. 3 shows the effect of 2-deoxy-D-glucose on IL-6, IL-4, TNF-alpha and IFN-gamma in serum of mice with chronic eczema.

Detailed Description

FIG. 1 shows the chemical structure of 2-deoxy-D-glucose. The invention provides a new application of 2-deoxy-D-glucose (2-DG), which is used for preparing a medicament for treating chronic eczema and is capable of adjusting the balance between auxiliary T cells Th1 and Th2, inhibiting the generation of inflammatory factors (TNF-alpha, INF-gamma and IL-6) and reducing ear swelling of a mouse model with chronic eczema.

The invention provides a medicament for treating chronic eczema, which comprises 2-deoxy-D-glucose with effective treatment amount and a medicinal carrier or excipient.

2-deoxy-D-glucose can be independently administered in the treatment of chronic eczema, or can be added with pharmaceutic adjuvant, or 2-deoxy-D-glucose and other medicines are combined and added with adjuvant to prepare various preparation formulations; the prepared dosage form can be oral dosage forms such as ointment, gel and the like, and can also be oral dosage forms such as tablets, capsules and the like.

The present invention is described in further detail below in connection with animal experiments, but is not meant to be limiting in any way.

Examples

Preparation of 2-deoxy-D-glucose ointment and blank ointment:

(1) taking 110g of stearic acid and 25g of glyceryl monostearate, mixing and heating to 85 ℃, and taking the mixture as an oily matrix; 9g of potassium carbonate, 3ml of triethanolamine, 100g of glycerol, 706ml of water and 40g of 2-deoxy-D-glucose powder are weighed, mixed and heated to 85 ℃ to be used as an aqueous substrate. After the two phases are completely dissolved, the aqueous base material is quickly transferred into the oily base material and continuously stirred, and the 4 percent 2-deoxy-D-glucose ointment is obtained after natural cooling.

(2) 2%, 0.5% 2-deoxy-D-glucose ointment and blank ointment preparation: the preparation method is the same as the above method by adjusting the addition amount of 2-deoxy-D-glucose powder to 20g, 5g and 0g respectively based on the above formula and adjusting the amount of water (726ml, 741ml and 746 ml).

Materials and methods

Experimental ICR mice, male and female halves, body weight (20. + -.2 g), were provided by Shanghai Spiker laboratory animal center.

Medicaments and agents

2-deoxy-D-glucose (purity: > 99%) is provided by Shanghai leaf biotechnology company, and compound dexamethasone acetate ointment is produced by Huarun Sanjiu medicine GmbH; dinitrochlorobenzene (DNCB), a product of shanghai chemical agents company, is prepared into 0.5% and 7% (mass ratio) dinitrochlorobenzene acetone solutions with acetone; enzyme-linked immunosorbent assay (ELISA) kit for IL-4, IL-6, TNF-alpha and IFN-gamma is purchased from Jiangsu enzyme immunoassay.

Preparation of model groups

The model group is the preparation of a mouse chronic eczema model: an ICR mouse is unhaired on the abdomen 1 day before the experiment, the area is about 2cm multiplied by 2cm, the abdomen of the mouse is coated with 100 microliter of 7 percent dinitro-chlorobenzene acetone solution for sensitization on the same day of the experiment, the abdomen of the mouse is coated with 100 microliter of 7 percent dinitro-chlorobenzene acetone solution again for secondary sensitization after 5 days, 20 microliter of 0.5 percent dinitro-chlorobenzene acetone solution is coated on the inner side of the right ear of the mouse for excitation on the second day after the secondary sensitization, and the left ear is coated with acetone solution for excitation 1 time every 3 days for 4 times.

Experimental grouping and administration

ICR mice were randomly divided into 6 groups of 8 mice per group, half male and female: a normal control group (blank ointment), a model group, a positive drug control group (compound dexamethasone acetate ointment) and a 2-deoxy-D-glucose ointment group (4%, 2%, 0.5%, g/g). The positive drug control group is coated with compound dexamethasone acetate ointment on the inner side of the right ear on the excitation day; the 2-deoxy-D-glucose ointment group was administered to the inner side of the right ear on the day of stimulation with 2-deoxy-D-glucose ointments at different concentrations; the normal control and model groups were coated with a blank ointment base without 2-deoxy-D-glucose. The drug was changed every day, and the mice were sacrificed 24 hours after the last challenge, and various indices were examined.

Observation index

1. Difference in ear weight: the experimental animals were sacrificed 24 hours after the last stimulation, a metal punch with a diameter of 0.9cm was used to punch holes in the middle of the ears, each tissue mass was weighed with an electronic balance, the difference in the weight of the left and right ears of each mouse was calculated, and the right ear pieces were made into paraffin blocks for use.

HE staining and observation: the paraffin blocks were cut into 5 μm thick sections, HE stained, and inflammatory cell infiltration was observed under a light microscope.

3. Serum IL-6, IL-4, TNF-alpha and IFN-gamma values: 1ml of blood is taken from a mouse through eyeballs, the mouse is centrifuged for 20 minutes at 1500r/min, serum is taken out and placed in a microfuge tube, the microfuge tube is stored at minus 80 ℃ for standby, an ELISA kit is adopted to measure IL-6, IL-4, TNF-alpha and IFN-gamma values in the serum, and the steps are carried out according to the kit specification.

Statistical treatment the assay results are expressed as x ± s, and differences between groups were performed using one-way anova.

Results

Effect of 2-deoxy-D-glucose on mouse model of Chronic eczema

(1) Difference in ear weight: compared with a normal control group, the right ear of the model group mice is obviously swollen, the weight of the model group mice is obviously increased compared with that of the left ear (P is less than 0.01), and the swelling condition of the right ear of the chronic eczema mice can be improved by the 2-deoxy-D-glucose ointment (4%, 2%, 0.5%) group and the compound dexamethasone acetate ointment group. The 2-deoxy-D-glucose ointment (4%) group and the 2-deoxy-D-glucose ointment (2%) group have significant difference (P <0.01) compared with the model group, and have dose-dependent relationship (Table 1, FIG. 2).

FIG. 2 is a graph showing the effect of 2-deoxy-D-glucose (2-DG) on the difference in ear weights of chronic eczema mice. Compared with the model group (20.7 +/-4.5 mg), the ear swelling degree (11.0 +/-4.4 mg) of the positive control group (DEX) is obviously reduced (P is less than 0.01), and the ear swelling degree (9.7 +/-5.6 mg) of the 2-deoxy-D-glucose ointment (4%) group is obviously reduced compared with the ear swelling degree (12.3 +/-5.18 mg) of the 2-deoxy-D-glucose ointment (2%) group (P is less than 0.01).

Table 1: effect of 2-deoxy-D-glucose on poor left and right ear weights of chronic eczema mice

Note: p <0.01 compared to normal control group; # P <0.01 compared to model group.

(2) Effect of 2-deoxy-D-glucose on the levels of IL-6, IL-4, TNF- α, IFN- γ in serum of Chronic eczema mice: at present, the inflammatory reaction of eczema caused by the interaction of internal and external factors is considered to be type IV hypersensitivity reaction similar to allergic contact dermatitis. Type iv hypersensitivity is an immune response mediated by T cells, activated by recognition of antigens by effector Th1 cells, releasing a variety of cytokines and chemokines. Wherein the tumor necrosis factor alpha (TNF-alpha) increases the expression of local vascular endothelial cell adhesion molecules, promotes the aggregation of mononuclear cells to the antigen existing part, and causes tissue damage; interferon gamma (IFN-gamma) and TNF-alpha can activate macrophage, further release proinflammatory cytokines IL-1 and IL-6 and the like to aggravate inflammatory response. In addition, IFN-gamma can inhibit proliferation of Th2 cells and inhibit IL-4 production by Th2 cells. IL-4 has the function of inhibiting Th1 cells and their functions, so IFN-gamma and IL-4 antagonize each other and maintain the functional balance of Th1/Th 2.

The experiment shows that compared with a normal control group, the serum IL-4 level of the mouse in the model group is reduced, and the IL-6, TNF-alpha and IFN-gamma levels are increased. Compared with a model group, the compound dexamethasone acetate ointment group can obviously increase the IL-4 level (P <0.01), reduce the IL-6, TNF-alpha and IFN-gamma levels (P <0.01), and the 2-deoxy-D-glucose ointment group with different concentrations increases with the administration concentration, the serum IL-4 level of the mouse increases (P <0.01), while the IL-6, TNF-alpha and IFN-gamma levels decrease (P <0.01) with the administration concentration (Table 2, figure 3), thereby regulating the functional balance of Th1/Th2 and inhibiting inflammatory reaction.

Table 2: effect of 2-deoxy-D-glucose on serum inflammatory factor of chronic eczema mouse

Effect of 2-deoxy-D-glucose on changes in otopathology in chronic eczematous mice: the ear tissue epidermal layer and the dermis layer of the normal control group mouse have regular structures and no inflammatory cells and congestion conditions; the horny layer of the ear tissue of the model group mouse is obviously thickened and cornified. Intercellular edema is accompanied by massive neutrophil infiltration. The compound dexamethasone acetate ointment and the 2-deoxy-D-glucose ointment in a high-dose group can obviously reduce the thickening of ear epidermis and inflammatory cell infiltration, and the prepared ointment containing the 2-deoxy-D-glucose can effectively treat chronic eczema.

The above embodiments are intended to illustrate rather than to limit the invention, and any modifications and variations of the present invention are within the spirit of the invention and the scope of the claims.

Claims

1. The application of 2-deoxy-D-glucose in preparing a medicament for treating chronic eczema is characterized in that the medicament contains effective dose of 2-deoxy-D-glucose and a medicinal carrier.