CN109700801B - A pharmaceutical composition for the treatment of psoriasis - Google Patents

Abstract

The invention designs a pharmaceutical composition for treating psoriasis, which consists of effective components and medically acceptable auxiliary materials, and is characterized in that the effective components consist of the following chemical components in percentage by weight: 45% -55% of aesculetin; 45-55% of betulinic acid. The pharmaceutical composition provided by the invention can obviously improve the dorsal skin PASI score of a psoriasis model mouse, reduce the epidermal thickening and weight loss of the mouse, and increase the proportion of regulatory T cells (Tregs) in the mouse body, and has a remarkable effect of treating psoriasis.

Description

A pharmaceutical composition for the treatment of psoriasis

Technical Field

The invention relates to medical preparations, in particular to a medicament containing organic compounds, which can be used for treating psoriasis.

Background

Psoriasis, also known as psoriasis, is one of the most common chronic inflammatory skin diseases, has stubborn disease conditions and is easy to recur, and can cause damage to the systemic system, and the main histopathological changes of the psoriasis are abnormal keratinocyte hyperplasia, parakeratosis, inflammatory cell infiltration, dermal tissue vascular hyperplasia and the like. Meanwhile, psoriasis is also easy to cause various complications such as psoriatic arthritis, metabolic syndrome, malignant tumor and the like, secondary damage is brought to patients, and the life quality and even the life cycle of the patients are seriously affected. However, the exact pathogenesis of psoriasis is not yet clear. Research has shown that the occurrence of psoriasis is related to a variety of factors including genetic, immunological, environmental influences, etc., with immune mediation being its currently generally accepted pathogenesis.

At present, the medicines for treating psoriasis mainly comprise antitumor medicines, calcipotriol, glucocorticoid, immunosuppressant, biological agents and the like. The medicines can play a certain treatment role clinically, but after long-term administration, side effects are generated, drug resistance and dependence are easy to form, and biological agents are expensive, so that huge economic burden is brought to patients. Therefore, researchers are actively searching for a new and effective anti-psoriasis drug in order to reduce side effects and burden on patients and society.

The traditional Chinese medicine has unique curative effect and advantage in the aspect of treating a plurality of immunological diseases. In recent years, many traditional Chinese medicine monomers have been successfully applied to the prevention and treatment of immune diseases, and have achieved remarkable achievements and promising development prospects. The learner Jie et al reported that camptothecin ointment has efficacy in treating psoriasis (Jie, Cinnamomum japonicum, Song Wen Rong, Shouchun. camptothecin ointment treats psoriasis 47 cases [ J ]. Chinese dermatology journal, Vol. 32, No. 2: 134-; in addition, the study of Zhao Dan et al reported that the effect of using cortex Fraxini bath in combination with narrow spectrum medium wave ultraviolet rays to treat psoriasis vulgaris was clear (Zhao Dan, Huangjianwei, Huangrui Reley. the clinical effect of using cortex Fraxini bath in combination with narrow spectrum medium wave ultraviolet rays to treat psoriasis vulgaris was observed [ J ]. the clinical journal of traditional Chinese medicine, Vol.27, No. 5 in 5.2015: 695 + 697). However, camptothecin is a class of alkaloids, which consists of hydroxycamptothecin, deoxycamptothecin, camptothecin, betulinic acid (Betulic acid), vincoside and the like, and it can be determined which specific camptothecin or camptothecins have biological activity for treating psoriasis; cortex fraxini mainly contains chemical components such as coumarins, lignans, seco-enol ether terpenoids, phenylethanoid glycosides, flavonoids, phenolic acids and triterpenes (Neonian, Linshijian, Zhang Bing. cortex fraxini chemical components and pharmacological action research progress [ J ] Chinese herbal medicine, volume 47, 18, 3332-3341 of 2016 year 9), which one or more chemical components can be used for treating psoriasis.

The invention content is as follows:

the invention aims to solve the technical problem of providing the pharmaceutical composition for treating psoriasis, and the pharmaceutical composition has obvious effect of treating psoriasis.

The technical scheme for solving the problems is as follows:

the pharmaceutical composition for treating psoriasis comprises effective components and medically acceptable auxiliary materials, and is characterized in that the effective components comprise the following chemical components in percentage by weight: 45% -55% of aesculetin; 45-55% of betulinic acid.

In the scheme, the optimal weight ratio of the chemical components is 50% of aesculetin and 50% of betulinic acid.

In the scheme, the molecular formula of the aesculetin is C₉H₆O₄Molecular weight of 178.14, and its chemical structure formula

The molecular formula of betulinic acid is C₃₀H₄₈O₃Molecular weight of 456.71, and its chemical structure formula

The pharmaceutical composition can be prepared into various conventional solid dosage forms, such as granules, tablets or capsules.

The solid dosage form is prepared by the following method:

weighing aesculetin and betulinic acid according to the prescription amount, mixing uniformly, adding corresponding auxiliary materials, and preparing into granules, tablets or capsules according to a conventional method.

The aesculetin and betulinic acid which form the effective components in the pharmaceutical composition have a synergistic effect, and the effect of treating psoriasis is remarkable.

The effective components in the pharmaceutical composition only consist of two compounds of aesculetin and betulinic acid, so that the pharmaceutical composition is easy to prepare and is beneficial to quality control.

To facilitate a better understanding of the present invention to the public, the following further illustrate the advantageous effects of the present invention through experiments on pharmaceutical effects and specific embodiments.

First, drug effect test

1. Laboratory animal

SPF grade BALB/C mice, body weight: 20 ± 2g, week old: male, free diet, temperature 25 ± 3 ℃, was maintained in SPF grade animal house, provided by experimental animals center, guangdong province, for 6 weeks.

2. Grouping animals

50 BALB/C mice were randomly divided into 7 groups, namely a normal group, a model group, an experimental group A, an experimental group B, an experimental group C and a control group 1-2, and 10 mice in each group.

3. Establishment of psoriasis model of mouse

The hair on the back of each mouse was shaved by electric hair clippers, and the remaining hair on the back of the mouse was removed by applying a depilatory cream to expose a skin area of about 3cm × 2 cm. Two days later, each mouse was applied with 62.5mg of 5% imiquimod ointment to the back skin daily for 7 consecutive days. The observation shows that the skin on the back of the mouse has obvious erythema, scale and thickening, namely the modeling is successful. Normal control mice were not given imiquimod ointment after back dehairing.

4. Test drugs and dosages

4.1 test drugs

Experimental group a: granules according to example 1 below; experimental group B: the tablet of example 2 below; experimental group C: the capsules of example 3 below;

control group 1: preparing 20g of aesculetin into granules according to the method in the application example 1;

control group 2: taking 20g of betulinic acid, and preparing into granules according to the method of the application example 1;

4.2 dosage administration

The test drugs of the experimental groups A-C and the control groups 1-2 were prepared into a reagent having a concentration equivalent to 5mg of the corresponding chemical component per ml using a physiological saline containing 10% (v/v) polyethylene glycol 400, respectively, and administered by gavage at a dose of 25mg/kg by weight of the corresponding chemical component 1 time a day for 7 days.

5. Experimental methods

5.1 mice dorsal skin PASI score

The skin condition of the back of the mice was recorded daily using a digital camera and scored according to the PASI scoring criteria. The scoring items included erythema (erythema), scales (scales) and infiltration thickening (strickness) of the mouse skin lesions. Each was classified by severity into 0-4 points, and the PASI scoring criteria were as follows: 0-none; 1-mild; 2-moderate; 3-severe; 4-very severe. The scores of each mouse and the total scores of the three mice in each group are averaged and compared.

5.2 mouse weight determination

The weight of the mice is weighed by a small-scale weighing machine every day, and compared with the weight of the mice before modeling and administration treatment, the daily change condition of the weight of the mice is calculated.

5.3 determination of epidermal thickness of mouse skin

After 7 days of molding and administration, animals were sacrificed, and mouse skin was fixed in 4% neutral formaldehyde, soaked overnight, dehydrated in a programmed dehydrator, embedded in a paraffin embedding machine, and then the paraffin-embedded tissue was cut into sections of 3 μm. Baking in a 65 deg.C oven for 1-2 hr, dewaxing to water, dyeing with hematoxylin for 10min, washing with tap water, returning blue, dyeing with eosin hydrochloride for 15min, dehydrating, sealing with neutral gum, air drying overnight, observing with microscope, taking picture, and calculating skin epidermis thickness.

5.4 Treg cell ratio determination in spleen and lymph node of mouse

After sacrifice, the spleen and lymph nodes were removed, ground through a 40 mesh screen to make a cell suspension, centrifuged to remove the supernatant, and the spleen cells were treated with lysed erythrocytes and washed 2 times with PBS along with the lymph node cells. 10 samples were taken after cell counting⁶The cells were resuspended in 100. mu.l PBS and 0.5. mu.l anti-CD4-FITC was added per sample and incubated for 30min at room temperature in the dark. Subsequently, the permeabilized cells are fixed, 0.5 mu l of anti-Foxp3-APC is added to each sample and incubated for 30min at room temperature in the dark, after PBS is washed once, 300 mu l of PBS is used for resuspending the cells, and the proportion of Treg cells is analyzed by a flow cytometer.

6. Statistical treatment

All data were statistically analyzed using SPSS V20 statistical software, and the data were measured using means. + -. standard deviation

And (4) showing.

7. Results of the experiment

7.1 mouse skin PASI score

Table 1 effect of pharmaceutical compositions on PASI score of dorsal skin of mice (n ═ 10)

Note: in comparison with the set of models,^*P＜0.05，^**p is less than 0.01; as compared with the control group 1, the control group,^#p is less than 0.05; comparison with control group 2⁺P＜0.05

The results in Table 1 show that the psoriasis model mice had a significant decrease in the dorsal skin PASI score (P < 0.05 or P < 0.01) following drug treatment. The mice of experimental groups A, B and C had significantly lower skin PASI than control groups 1 and 2, indicating that the pharmaceutical composition had a better improvement in the dorsal skin of the mice (P < 0.05) than the existing control drugs.

7.2 mouse weight changes

Table 2 effect of pharmaceutical composition on weight change of mice (n ═ 10)

Note: in comparison with the set of models,^*P＜0.05，^**p is less than 0.01; as compared with the control group 1, the control group,^#p is less than 0.05; comparison with control group 2⁺P＜0.05

The results in Table 2 show that the psoriasis model mice, after being treated with the drug, have a significant and reduced weight loss (P < 0.01). Compared with the control groups 1 and 2, the weight loss of the mice of the experimental groups A, B and C is remarkably relieved, and the pharmaceutical composition has better improvement effect (P is less than 0.05) on the weight loss of the mice compared with the existing control drugs.

7.3 thickness of epidermal layer of mouse skin

Table 3 effect of pharmaceutical composition on thickness of skin epidermis in mouse (n ═ 10)

Note: in comparison with the set of models,^*P＜0.05，^**p is less than 0.01; as compared with the control group 1, the control group,^##p is less than 0.01; comparison with control group 2⁺⁺P＜0.01

The results in Table 3 show that the epidermal thickness is significantly reduced (P < 0.01) after the drug treatment in psoriasis model mice. Compared with the control groups 1 and 2, the thickness of the mouse epidermis of the experimental group A, B and the mouse epidermis of the experimental group C are obviously reduced, which shows that compared with the existing control drug, the drug composition has better improvement effect on the thickening condition of the mouse epidermis (P is less than 0.01).

7.4Treg cell ratio

Table 4 effect of pharmaceutical compositions on Treg cells in mice (n ═ 10)

Note: in comparison with the set of models,^*P＜0.05，^**p is less than 0.01; as compared with the control group 1, the control group,^#p is less than 0.05; comparison with control group 2⁺P＜0.05

The results in Table 4 show that the proportion of Treg cells in both spleen and lymph nodes was significantly increased (P < 0.05 or P < 0.01) after drug treatment in psoriasis model mice. Compared with the control groups 1 and 2, the proportion of Treg cells in the spleen and lymph nodes of the mice of the experimental groups A, B and C is remarkably increased, and the fact that compared with the existing control drug, the pharmaceutical composition has a better effect of improving the Treg cells in the mice (P is less than 0.05) is shown.

8. Conclusion

The experimental result shows that compared with the single use of the aesculetin and the betulinic acid, the pharmaceutical composition has a synergistic effect. Meanwhile, compared with aesculetin and betulinic acid in different weight ratios, the medicinal composition has a better treatment effect when the weight ratio is 50% to 50%, and the result shows that the medicinal composition can effectively treat psoriasis.

Detailed Description

Example 1

Weighing 20g of aesculetin and 20g of betulinic acid according to the weight, uniformly mixing, adding 80g of powdered sugar, 52g of dextrin, 48g of lactose and a proper amount of 65% ethanol to prepare a soft material, sieving to prepare granules, drying at 60 ℃ under normal pressure, finishing granules, and packaging into granules by adopting a packaging device, wherein each bag is 1 g. The granule is administered orally 1 bag each time, 1 time per day, and 7 days as a treatment course, and can be continuously administered for 2-3 treatment courses.

Example 2

Weighing 22g of aesculetin and 18g of betulinic acid according to the weight, uniformly mixing, adding 80g of powdered sugar, 52g of dextrin, 48g of lactose and a proper amount of 65% ethanol to prepare a soft material, sieving to prepare granules, drying at 60 ℃ under normal pressure, adding 6.6g of magnesium stearate with the particle amount of 3% after finishing granules, uniformly mixing, and pressing the granules into 440 tablets by using a punching device. The tablet is administered orally, 2 tablets each time, 1 time per day, and 7 days as a treatment course, and can be continuously administered for 2-3 treatment courses.

Example 3

Weighing 18g of aesculetin and 22g of betulinic acid according to the weight, uniformly mixing, adding 80g of powdered sugar, 52g of dextrin, 48g of lactose and a proper amount of 65% ethanol to prepare a soft material, sieving to prepare granules, drying at 60 ℃ under normal pressure, finishing granules, and filling into No. 3 empty capsules, wherein the average content of each capsule is 500 mg. The capsule is administered orally 2 granules each time, 1 time per day, and 7 days as a treatment course, and can be continuously administered for 2-3 treatment courses.

Claims (4)

1. The pharmaceutical composition for treating psoriasis comprises effective components and medically acceptable auxiliary materials, and is characterized in that the effective components comprise the following chemical components in percentage by weight: 45% -55% of aesculetin and 45% -55% of betulinic acid.

2. The pharmaceutical composition for treating psoriasis according to claim 1, wherein the effective components consist of the following chemical components in percentage by weight: 50% of aesculetin and 50% of betulinic acid.

3. The pharmaceutical composition for treating psoriasis according to claim 1 or 2, wherein the pharmaceutical composition is in the form of granules, tablets or capsules.

4. The pharmaceutical composition for treating psoriasis according to claim 3, which is prepared by the following method:

weighing aesculetin and betulinic acid according to the prescription amount, mixing uniformly, adding corresponding auxiliary materials, and preparing into granules, tablets or capsules according to a conventional method.