Pharmaceutical composition for chronic skin inflammation

Abstract

The invention discloses a pharmaceutical composition for chronic scytitis, and belongs to the technical field of pharmaceutical compositions. The pharmaceutical composition for scytitis comprises an ethanol solution of a tripterygium wilfordii extract with the concentration of 15 mg / mL and a hydrocortisone solution with the mass percentage content of 1%. The combination of the tripterygium wilfordii extract and the hydrocortisone in the pharmaceutical composition for skin inflammation has a good synergistic anti-inflammatory effect, and side effects of the hydrocortisone are reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical compositions, in particular to a pharmaceutical composition for treating chronic skin inflammation. Background Art

[0002] Atopic dermatitis (AD) is a common, chronic, relapsing inflammatory skin disease that often lasts a lifetime and severely impacts patients' quality of life. The pathogenesis of AD is complex, involving the interplay of multiple factors, including genetic susceptibility, skin barrier dysfunction, dysbiosis of the skin microbiome, and immune dysfunction. These factors collectively lead to persistent and exacerbated skin inflammation, triggering typical AD symptoms such as intense itching, erythema, papules, and dry skin.

[0003] Currently, the treatment of AD mainly relies on glucocorticoids, such as hydrocortisone ointment (2.5%). This type of drug quickly controls the inflammatory response of AD by inhibiting the accumulation of inflammatory cells (such as macrophages and leukocytes) at the site of inflammation, as well as inhibiting phagocytosis, the release of lysosomal enzymes, and the synthesis and release of inflammatory chemical mediators. However, although glucocorticoids have significant anti-inflammatory effects in the short term, their long-term use may lead to a series of adverse reactions, such as skin atrophy, capillary dilation, pigmentation, and secondary infections, and may even cause systemic side effects, such as adrenal cortex function suppression. Therefore, the use of glucocorticoids should be cautious and should not be applied over a large area for a long time. Therefore, we urgently need an alternative, long-term dosing regimen.

[0004] Among traditional Chinese medicinal herbs, Tripterygium wilfordii has attracted considerable attention for its remarkable effects in dispelling wind and dampness, and in promoting blood circulation and unblocking meridians. Its bitter and cold nature has a powerful heat-clearing effect, particularly in reducing swelling and alleviating pain, and it has unique therapeutic effects on conditions such as heat arthritis and stubborn arthritis. Modern pharmacological studies have shown that Tripterygium wilfordii has broad application value in the treatment of various inflammatory skin diseases, including Behçet's disease, eczema, psoriasis, leprosy, scabies, and stubborn ringworm. Currently, preparations such as triptolide ointment (10g: 200μg) are used to treat skin inflammation. However, long-term, high-dose topical application of Tripterygium wilfordii preparations may be absorbed through the skin into the bloodstream, causing systemic side effects such as liver and kidney damage, limiting their clinical application.

[0005] In summary, although existing glucocorticoids and tripterygium wilfordii preparations have some efficacy in the treatment of AD, their long-term safety and tolerability remain significant issues. Therefore, there is an urgent need to develop safer and alternative treatment options to improve the long-term treatment outcomes and quality of life of AD patients. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to propose a pharmaceutical composition for chronic skin inflammation. The pharmaceutical composition optimizes the dosage ratio of hydrocortisone solution and tripterygium wilfordii extract, aims to reduce adverse reactions, improve the therapeutic effect while reducing the dosage of both, and provide patients with skin inflammation with a safer and more effective treatment option.

[0007] The pharmaceutical composition of the hydrocortisone solution and the tripterygium wilfordii extract of the present invention can effectively treat AD and reduce the side effects of the hydrocortisone solution and the tripterygium wilfordii extract.

[0008] The purpose of the present invention is achieved through the following technical solutions:

[0009] A pharmaceutical composition for chronic skin inflammation, comprising a tripterygium wilfordii extract and hydrocortisone; further, the skin inflammation is atopic dermatitis.

[0010] Preferably, the Tripterygium wilfordii extract is an ethanol solution of the Tripterygium wilfordii extract with a mass concentration of 15 mg / mL; and the hydrocortisone is a hydrocortisone solution with a mass percentage of 1%.

[0011] The preparation method of the Tripterygium wilfordii extract comprises the following steps:

[0012] Take the Chinese medicinal material of Tripterygium wilfordii, grind it, and add anhydrous ethanol to soak overnight. Preferably, the mass volume ratio (m / V) of Tripterygium wilfordii to anhydrous ethanol is 1:80-120, preferably 1:100, such as 1 mg of Tripterygium wilfordii is added to 100 mL of anhydrous ethanol;

[0013] After filtering out the medicinal residue, the anhydrous ethanol solution is concentrated by rotary evaporation into a brown liquid, which is then evaporated in a water bath at 60°C and finally dried to obtain a dark brown tripterygium wilfordii extract; the tripterygium wilfordii extract is dissolved in anhydrous ethanol to prepare tripterygium wilfordii extract solutions of different concentrations for use.

[0014] The pharmaceutical composition of the present invention comprises a tripterygium wilfordii extract solution with a mass concentration of 15 mg / mL and a hydrocortisone solution with a mass percentage of 1%.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] (1) The combination of the two drugs has a good synergistic anti-inflammatory effect: The present invention optimizes the usage ratio of hydrocortisone solution and tripterygium wilfordii extract, so that hydrocortisone and tripterygium wilfordii extract can synergistically exert a stronger anti-inflammatory effect, can quickly control the inflammatory response, and show a stronger inhibitory effect on the expression of inflammation-related genes when the two drugs are used in combination.

[0017] (2) Reducing the side effects of glucocorticoids (hydrocortisone): The present invention reduces the dosage of hydrocortisone by introducing Tripterygium wilfordii extract, and weakens the adverse reactions caused by long-term and excessive use of glucocorticoids (hydrocortisone), such as skin atrophy, thinning, and capillary dilation. The expression of genes related to proliferation (PCNA, Cyclin-D1), apoptosis (Stp1), lipid synthesis (HMG-CoA), and filaggrin (Fillagrin) in mouse skin was examined by qPCR technology. It was found that Tripterygium wilfordii extract can effectively reverse the inhibition of hydrocortisone on the expression of various genes. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is the experimental flow chart of Example 2;

[0019] Figure 2 Photos of mice in the eight experimental groups in Example 2 and HE-stained photos of ear tissues of the corresponding groups;

[0020] Figure 3 Graph showing ear thickness changes over time for eight groups of mice in Example 2;

[0021] Figure 4 is a graph of the ear weights of mice in the eight groups in Example 2;

[0022] Figure 5 This is a graph showing the expression of genes related to ear inflammation in mice in the normal control group, model group, treatment groups 1-3, and positive control group in Example 2;

[0023] Figure 6 Graph showing the expression of genes related to proliferation, apoptosis, lipid synthesis, and filaggrin in the skin of mice in the normal control group, treatment groups 1-3, and treatment group 5 in Example 3;

[0024] Figure 7 Graph showing the expression of mouse dermis-related genes (COL1A1, COL3A1, MMP3, MMP9) in the normal control group, treatment groups 1-3, and treatment group 5 in Example 3. DETAILED DESCRIPTION

[0025] In order to enable those skilled in the art to understand the present invention more clearly and intuitively, the present invention will be further described below with reference to the accompanying drawings.

[0026] Example 1

[0027] Preparation of Tripterygium wilfordii extract

[0028] 30g of Tripterygium wilfordii root was crushed and soaked overnight in 300mL of anhydrous ethanol. After filtering to remove the residue, the 300mL anhydrous ethanol solution was rotary evaporated to approximately 25mL of a brown liquid. The entire ethanol solution was then heated in a water bath at 60°C for evaporation. Finally, the solution was dried to obtain a dark brown solid, which was the Tripterygium wilfordii extract.

[0029] When in use, the solid is weighed and prepared with anhydrous ethanol to obtain a solution of the Tripterygium wilfordii extract with an ideal concentration.

[0030] Preparation of hydrocortisone solution

[0031] Take 3.62g of hydrocortisone, dissolve it in 100ml of propylene glycol-ethanol mixed solution (50:50), and stir to mix.

[0032] Hydrocortisone, also known as cortisol, is an organic compound with the chemical formula C 21 H 30 O5, extracted from the adrenal cortex, is a type of glucocorticoid and has the strongest effect on carbohydrate metabolism. Appearance: White or off-white crystalline powder, insoluble in water, poorly soluble in ether, slightly soluble in chloroform, and soluble in acetone and ethanol.

[0033] Example 2

[0034] Animal experiments on the treatment of acute AD, comparing the therapeutic effects of drug compositions with different ratios

[0035] In this example, the widely used phorbol 12-myristate 13-acetate (PMA)-induced acute / chronic skin inflammation model was used to study the therapeutic effect of drugs on atopic dermatitis (AD).

[0036] C57 mice were divided into 8 groups (normal control group, model group, treatment groups 1-5, and positive treatment group), with 5 mice in each group, for a total of 40 mice:

[0037] ①Normal control group: no sensitization;

[0038] ②Model group: sensitization;

[0039] ③ Treatment group 1: sensitization + hydrocortisone solution (1%);

[0040] ④ Treatment group 2: sensitization + Tripterygium wilfordii extract solution (15 mg / ml);

[0041] ⑤ Treatment group 3: sensitization + Tripterygium wilfordii extract solution (15 mg / ml) + hydrocortisone solution (1%) combination;

[0042] ⑥ Treatment group 4: sensitization + Tripterygium wilfordii extract solution (30 mg / ml);

[0043] ⑦ Treatment group 5: sensitization + Tripterygium wilfordii extract solution (30 mg / ml) + hydrocortisone solution (1%) combination;

[0044] ⑧Positive treatment group: sensitization + hydrocortisone solution (2.0%).

[0045] Except for the normal control group, 10 μL of 0.25% phorbol ester (PMA) solution was evenly applied to the inside and outside of the right ears of the mice in the model group, treatment groups 1-5, and positive treatment group for sensitization. 30 minutes after sensitization, the right ears of the mice were medicated, and 20 μl of treatment solution was administered inside and outside each treatment group, twice a day. Among them, the combination medication group mixed the Tripterygium wilfordii extract solution and hydrocortisone solution to form a mixed solution and then smeared it for medication. 1h, 2h, 4h, 8h, 12h, 24h and 48h after the first medication, the ears of the mice in each group were observed and recorded, and the thickness of the ears (0.01mm) of each time point was measured with a thickness gauge, and data were recorded and calculated and analyzed. Subsequently, the mice were killed, their ears were taken, and the ears were weighed to examine the weight differences of the ears of the mice in each group; the ear tissues were HE stained and observed. The flow chart is as follows Figure 1 shown.

[0046] The head photos of mice in each group and the corresponding HE staining results of ear tissues are shown in the figure. Figure 2 As shown. Figure 2 It can be seen that the inflammation of the ear tissue of mice in treatment group 1 and treatment group 2 was more serious, but after treatment with 15 mg / mL tripterygium wilfordii extract solution + 1% hydrocortisone solution, the inflammation was significantly improved, and the effect was close to the treatment effect of 30 mg / mL tripterygium wilfordii extract solution or 2% hydrocortisone solution.

[0047] Inflammation sites are always accompanied by "redness, swelling, heat and pain". The thickness of the mouse ears reflects the severity of inflammation. When the thickness of the mouse ears increases, it means that the inflammation on the mouse ears is more severe. Figure 3 As shown, from Figure 3 The results show that the ear thickness problem of mice in treatment groups 1 and 2 was improved very little, while in treatment group 3, the ear thickness problem was significantly improved after treatment with 15 mg / mL Tripterygium wilfordii extract solution + 1% hydrocortisone solution, indicating that the combined treatment of Tripterygium wilfordii extract solution and hydrocortisone solution produced a synergistic effect in reducing the thickness of mice's ears.

[0048] The ear weights of mice in each group were Figure 4 As shown, from Figure 4The results shown show that compared with the model group, the ear weight of mice in treatment groups 1 and 2 did not improve much, while in treatment group 3, the ear weight problem was significantly improved after treatment with 15 mg / mL Tripterygium wilfordii extract solution + 1% hydrocortisone solution, indicating that the combined treatment of Tripterygium wilfordii extract solution and hydrocortisone solution produced a synergistic effect on reducing the ear weight of mice.

[0049] Furthermore, the expression of inflammation-related genes (IL-1α, IL-1β, IL-6, IL-4, IL-13, TNF-α) was detected in the inflamed areas of the ears of mice in the normal control group, model group, treatment groups 1-3, and positive control group. The results are as follows Figure 5 As shown. Figure 5 The results showed that the treatment with 15 mg / mL Tripterygium wilfordii extract solution and 1% hydrocortisone solution produced a synergistic effect.

[0050] Example 3

[0051] In this example, different treatment regimens were used to administer the drug to normal animals to investigate and compare the side effects of the drug compositions at different ratios.

[0052] C57 mice were divided into 7 groups (blank control group, treatment groups 1-5, positive control group), 5 mice in each group, for a total of 35 mice:

[0053] ①Normal control group;

[0054] ② Treatment group 1: hydrocortisone solution (1%);

[0055] ③ Treatment group 2: Tripterygium wilfordii extract solution (15 mg / ml);

[0056] ④ Treatment group 3: combination of Tripterygium wilfordii extract solution (15 mg / ml) and hydrocortisone solution (1%);

[0057] ⑤ Treatment group 4: Tripterygium wilfordii extract solution (30 mg / ml);

[0058] ⑥ Treatment group 5: combination of Tripterygium wilfordii extract solution (30 mg / ml) and hydrocortisone solution (1%);

[0059] ⑦ Positive treatment group: hydrocortisone solution (2.0%).

[0060] Subsequently, the drug was applied to the skin of the mice for three consecutive days, twice a day. Among them, the combination drug group mixed the Tripterygium wilfordii extract solution and the hydrocortisone solution to form a mixed solution and then applied it. After the last administration, the skin and internal organs of the mice were collected. The expression of proliferation (PCNA, Cyclin-D1), apoptosis (Stp1), lipid synthesis (Hmg-CoA) and filamentous protein (Fillagrin) genes in the skin of mice in the normal control group, treatment groups 1-3, and treatment group 5 was examined by qPCR technology. The results are as follows Figure 6 As shown. Figure 6 The results of treatment groups 1 and 3 showed that long-term use of hydrocortisone solution (glucocorticoid) would reduce related expressions on the epidermis. Proliferation genes (PCNA, Cyclin-D1) reflect the regenerative ability of epidermal keratinocytes, a key process for barrier repair. The epidermis of AD patients is often damaged by inflammation and scratching and needs to be repaired quickly. The apoptosis gene (Stp1) reflects that glucocorticoids can inhibit keratinocyte apoptosis, resulting in slower epidermal cell renewal. The lipid synthesis gene (Hmg-CoA) is involved in epidermal cholesterol synthesis and affects the integrity of the skin barrier lipid layer. Hydrocortisone inhibits the expression of this gene and affects epidermal cholesterol synthesis. In addition, filaggrin is the end product of epidermal differentiation and maintains the skin's physical barrier and moisturizing function. The expression of the filaggrin gene is inhibited, but tripterygium wilfordii extract can effectively reverse the inhibition of hydrocortisone on the expression of various genes.

[0061] Furthermore, the expression of dermal collagenase genes (COL1A1, COL3A1) and matrix metalloproteinase genes (MMP3, MMP9) in the normal control group, treatment groups 1-3, and treatment group 5 was investigated by qPCR. Type I and type III collagen are the main structural proteins of the dermis, maintaining skin thickness and mechanical strength. In addition, matrix metalloproteinases are involved in tissue remodeling. The results are as follows. Figure 7 As shown in Figure 2, long-term use of hydrocortisone solution (glucocorticoid) significantly inhibits the expression of these genes. However, Tripterygium wilfordii extract can effectively reverse this inhibitory effect.

[0062] From the above, we can see that Tripterygium wilfordii extract can not only synergize with hydrocortisone to treat skin inflammation, but also reduce the negative effects of hydrocortisone on the skin, such as skin atrophy, skin thinning, capillary dilation and other problems.

Claims

1. A pharmaceutical composition for chronic skin inflammation, characterized in that: Includes Tripterygium wilfordii extract and hydrocortisone.

2. The pharmaceutical composition according to claim 1, wherein The invention relates to a mixed solution consisting of a tripterygium wilfordii extract solution and a hydrocortisone solution.

3. The pharmaceutical composition according to claim 1, wherein The Tripterygium wilfordii extract solution is an ethanol solution of the Tripterygium wilfordii extract with a mass concentration of 15 mg / mL.

4. The pharmaceutical composition according to claim 1, wherein The hydrocortisone is a hydrocortisone solution with a mass percentage of 1%.

5. The pharmaceutical composition according to claim 1, wherein The chronic skin inflammation is atopic dermatitis.

6. The pharmaceutical composition according to claim 1, wherein The preparation method of the Tripterygium wilfordii extract comprises the following steps: Crush the Tripterygium wilfordii Chinese medicinal material, add anhydrous ethanol and soak overnight; After filtering out the medicinal residue, the anhydrous ethanol solution was concentrated by rotary evaporation to form a brown liquid, which was then evaporated by heating in a water bath at 60° C. and finally dried to obtain a dark brown Tripterygium wilfordii extract.

7. The pharmaceutical composition according to claim 6, wherein The mass volume ratio of tripterygium wilfordii to anhydrous ethanol is 1:80-120.

8. The pharmaceutical composition according to claim 4, wherein The preparation method of hydrocortisone solution comprises the following steps: The hydrocortisone is dissolved in a propylene glycol-ethanol mixed solution, wherein the volume ratio of propylene glycol to ethanol in the propylene glycol-ethanol mixed solution is 1:

1.

9. Use of Tripterygium wilfordii extract and hydrocortisone in the preparation of a drug for treating atopic dermatitis.

10. Use of a tripterygium wilfordii extract in the preparation of a drug for reducing the side effects of hydrocortisone, the side effects of hydrocortisone including skin atrophy, skin thinning, and capillary dilation.

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