Lilac-free angelica and white peony extract, and preparation method and application thereof

Abstract

This invention discloses an extract of *Eupatorium fortunei* and *Angelica dahurica*, its preparation method, and its application, comprising the following steps: weighing *Eupatorium fortunei* and *Angelica dahurica* at a weight ratio of 1:1-2 and setting aside; mixing the raw materials from step (1) with water at a material-to-liquid ratio of 1:10-15 (g / mL) and heating, and extracting by decoction 2-3 times; performing solid-liquid separation after each extraction in step (2), collecting the filtrates from each extraction, and mixing them for later use; concentrating and drying the mixed filtrate from step (3) to obtain the finished dry powder. The prepared dry powder has significant effects in treating inflammatory skin diseases and repairing the skin barrier.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to an extract of Linglingxiang and Baizhi, its preparation method and application. Background Technology

[0002] Seborrheic dermatitis is a common chronic inflammatory skin disease characterized by erythema, oily or dry scaling or crusting, accompanied by itching. It commonly occurs in areas rich in sebaceous glands, such as the scalp, behind the ears, face (nasolabial folds, upper lip, eyelids, eyebrows), and upper chest. Although this disease is benign, non-contagious, and non-fatal, it is difficult to cure and prone to recurrence, gradually impacting people's quality of life, especially women, young people, and those with severe symptoms. This has made seborrheic dermatitis one of the key diseases for prevention and treatment worldwide. Its pathogenesis is closely related to the overactivation of the inflammatory response; the abnormal release of inflammatory mediators is a key trigger for skin erythema, itching, and desquamation.

[0003] Currently, clinical drug treatments are divided into two categories: topical and oral.

[0004] Topical treatment: Corticosteroids, nonsteroidal anti-inflammatory drugs (NSAIDs), antifungals, and calcineurin inhibitors are commonly used to treat seborrheic dermatitis. While these treatments offer significant short-term efficacy, long-term use can lead to side effects, drug resistance, relapse, and long-term dependence. According to a review of seborrheic dermatitis treatment, corticosteroids can cause side effects such as telangiectasia, hirsutism, atrophy, and perioral dermatitis; therefore, prolonged use is not recommended. Studies by Youn HJ et al. have shown that NSAIDs applied to the skin can cause stinging, itching, burning, and erythema.

[0005] Oral medications mainly include antihistamines, B vitamins, short-term low-dose glucocorticoids, and traditional Chinese medicine. However, oral medications may have potential effects on the gastrointestinal tract, liver and kidney function, and patients have poor tolerance to them, making it difficult to adhere to long-term treatment and seriously affecting their quality of life.

[0006] Therefore, developing safe, effective, and minimally invasive anti-inflammatory drugs has become an urgent need in the current treatment of seborrheic dermatitis. Summary of the Invention

[0007] In view of the shortcomings of the existing technology, the present invention provides an extract of Linglingxiang and Angelica dahurica, a preparation method thereof and its application, which has significant effects on anti-inflammatory treatment and skin barrier repair in the treatment of seborrheic dermatitis and dandruff.

[0008] To achieve the above objectives, the present invention provides a method for preparing extracts of *Eupatorium fortunei* and *Angelica dahurica*, comprising the following steps: (1) Weighing raw materials Weigh out Linglingxiang and Baizhi according to a weight ratio of 1:1-2, and set aside. (2) Extraction by decoction The raw materials in step (1) are mixed with water at a ratio of 1:10-15 (g / mL) and heated, and then extracted by decoction 2-3 times. (3) Filtration After each extraction in step (2), perform solid-liquid separation, collect the filtrates from each extraction, and mix them for later use. (4) Finished product preparation The mixed filtrate from step (3) is concentrated and dried to obtain the finished dry powder.

[0009] Secondly, the present invention also provides an extract prepared by the above-described preparation method.

[0010] Thirdly, this invention relates to the application of the above-mentioned extracts in the preparation of pharmaceuticals and topical products for treating inflammatory skin diseases and repairing the skin barrier. It shows particularly significant effects in anti-inflammatory treatment and skin barrier repair in treating seborrheic dermatitis and dandruff.

[0011] Compared with existing technologies, the present invention has the following beneficial effects: The present invention, through a specific combination of Lingling water extract and Angelica dahurica water extract, targets the core TCM pathogenesis of seborrheic dermatitis, namely internal damp-heat and external wind-evil invasion, to achieve the synergistic effect of resolving dampness and relieving itching and dispelling wind and drying dampness, thereby achieving the purpose of treating seborrheic dermatitis. Furthermore, animal experimental data have fully confirmed the outstanding technical effect of the combined use of the two in the treatment of seborrheic dermatitis, making the extract of the present invention have significant effects in the anti-inflammatory treatment of seborrheic dermatitis and dandruff, as well as in repairing the skin barrier. Attached Figure Description

[0012] Figure 1 These are comparative images of the skin appearance of four groups of guinea pigs after 14 days of drug administration in the toxicity verification experiment of this invention; Figure 2 This is a comparison graph of line graphs showing the changes in body weight (R) and food intake (I) of four groups of guinea pigs after 14 days of drug administration in the toxicity verification experiment of this invention. Figure 3 This is a comparative image of the skin tissue pathological characteristics of four groups of guinea pigs after 14 days of drug administration in the toxicity verification experiment of this invention; Figure 4 This is a comparison of skin lesion changes in 5 groups of guinea pigs at different time points in an experiment to verify the effectiveness of this invention. Figure 5 A comparative graph showing the changes in skin lesion scores (B) and body weight (C) of five groups of guinea pigs at different time points in an experiment to verify the effectiveness of this invention; Figure 6 This is a comparative image of the skin pathological characteristics of 5 groups of guinea pigs 14 days after drug administration in the efficacy verification experiment of this invention; Figure 7 The expression levels of inflammatory factors in the serum of four groups of guinea pigs were measured in an experiment to verify the effectiveness of this invention. Figure 8 The total ion chromatogram is obtained by ultra-high performance liquid chromatography-orbit trap high-resolution mass spectrometry detection of the extract powder in Example 1 of the present invention. Detailed Implementation

[0013] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments.

[0014] Currently, clinical anti-inflammatory treatment for seborrheic dermatitis mainly relies on chemical drugs such as corticosteroids, antifungal medications, and calcineurin inhibitors. However, long-term use of chemical drugs can easily lead to side effects such as hormone-dependent dermatitis, skin atrophy, and flora imbalance, and the condition is highly prone to relapse after discontinuation. Traditional Chinese medicine, on the other hand, requires specific treatments, including oral and / or topical administration, based on individual patient conditions, making it difficult to meet the core requirements of "safety and gentleness, stable anti-inflammatory efficacy, and reduced recurrence" in the clinical treatment of seborrheic dermatitis.

[0015] To address the aforementioned issues, this invention provides an extract that innovatively combines *Linglingxiang* and *Baizhi*. Compared to existing applications of single-herb extracts, this combined use is unconventional and effectively avoids the drawbacks of using a single extract (*Linglingxiang* or *Baizhi*).

[0016] The existing technology gaps in the use of Linglingxiang extract: The application scenarios of Linglingxiang in the existing technology are highly limited, only used as a food flavoring or as medicine for the treatment of systemic symptoms such as colds, headaches, and abdominal distension. Its disclosed anti-inflammatory effects are only for the relief of systemic inflammation. There have never been any reports of preparing it into an aqueous extract suitable for topical skin administration for the treatment of local inflammatory skin diseases such as seborrheic dermatitis. Furthermore, there is no existing technology that reveals that it can be used in combination with other Chinese herbal extracts to achieve synergistic enhancement of anti-inflammatory effects. It is completely unsuitable for the localized and precise anti-inflammatory treatment needs of seborrheic dermatitis.

[0017] Existing technological bottlenecks of Angelica dahurica extract: Although Angelica dahurica can be used as a food flavoring or as a medicine for some skin conditions such as chronic inflammatory pain, eczema, and acne, when applied to skin inflammation, it can only exert limited anti-inflammatory effects through a single component and a single pathway, and cannot effectively control the complex inflammatory network mediated by multiple pathways in seborrheic dermatitis. At the same time, existing technologies have not yet solved the potential photosensitivity and toxicity problem when Angelica dahurica is applied topically, resulting in insufficient safety for long-term skin administration and making it impossible to achieve long-term stable control of seborrheic dermatitis.

[0018] This invention utilizes a specific combination of Lingling water extract and Angelica dahurica water extract to target the core TCM pathogenesis of seborrheic dermatitis, which involves internal damp-heat and external wind invasion. This combination achieves a synergistic effect of resolving dampness and relieving itching, as well as dispelling wind and drying dampness, thereby achieving the goal of treating seborrheic dermatitis.

[0019] Specifically, the preparation methods and applications of extracts of *Linglingxiang* and *Baizhi* are as follows.

[0020] This invention provides a method for preparing extracts of *Eupatorium fortunei* and *Angelica dahurica*, comprising the following steps: (1) Weighing raw materials After being cleaned, dried, and pulverized, Linglingxiang and Baizhi were weighed out in a weight ratio of 1:1-2 and set aside for later use.

[0021] (2) Extraction by decoction Mix the raw materials and water in step (1) at a ratio of 1:10-15 (g / mL) and heat until boiling. Then maintain a gentle boil and stir every 5-10 minutes. Extract by decoction 2-3 times. Calculate the extraction time after boiling, and each extraction time is 30-50 minutes.

[0022] (3) Filtration After each extraction in step (2), vacuum filtration is used to separate the solid and liquid, and the filtrates from each extraction are collected and mixed for later use. (4) Finished product preparation After concentrating the mixed filtrate in step (3) to a relative density of 1.10-1.20 (25℃), it is dried to obtain the finished dry powder.

[0023] To verify the application of the above-mentioned extracts in treating inflammatory skin diseases and repairing the skin barrier, the present invention will provide a variety of specific extract products through the following multiple embodiments.

[0024] The raw material used in this invention is *Linglingxiang* (Latin name: *Linglingxiang*). Lysimachiafoenum-graecumHance ), Angelica dahurica (Latin name: Angelicadahurica Both are Chinese medicinal materials that meet the standards of the Pharmacopoeia of the People's Republic of China.

[0025] Example 1 An extract of *Eupatorium fortunei* and *Angelica dahurica*, prepared by the following method: 1. Raw material ratio Linglingxiang and Baizhi were cleaned, dried, and pulverized separately for later use; The weight ratio of Linglingxiang to Baizhi is 1:1. Weigh them out and set them aside.

[0026] 2. Extraction by decoction Mix the raw material (a mixture of coarse powder of Linglingxiang and Baizhi) with purified water at a material-to-liquid ratio of 1:10 (g / mL), place it in an electric heating mantle, heat to 95-100℃, and after boiling, reduce to low heat and maintain a gentle boil. Stir once every 5-10 minutes to ensure thorough extraction. Extract twice, each time for 30 minutes. 3. Filtration After each extraction, the solution was filtered using a filter flask and a vacuum pump. The filtrate was collected, and the two filtrates were combined to obtain the aqueous extract of Linglingxiang-Baizhi. 4. Concentration The crude extract was placed in a rotary evaporator and concentrated under a vacuum of 0.06–0.08 MPa and a temperature of 50–60 °C until the relative density of the concentrate was 1.10–1.20 (25 °C).

[0027] 5. Drying The concentrated medicinal liquid was continuously dried in a dryer to obtain a dry powder of Linglingxiang-Baizhi water extract, which was then sealed and stored in a desiccator for later use. The powder was then analyzed by ultra-high performance liquid chromatography-orbit trap high-resolution mass spectrometry (e.g., Figure 8 The dried medicinal powder is qualified.

[0028] Example 2 The difference between this embodiment and Embodiment 1 is that: In step (1), the weight ratio of Linglingxiang to Baizhi is 1:1.2; In step (2), the material-to-liquid ratio is 1:12 (g / mL), and the extraction time is 40 min.

[0029] Example 3 The difference between this embodiment and Embodiment 1 is that: In step (1), the weight ratio of Linglingxiang to Baizhi is 1:1.5; In step (2), the extraction time is 50 min.

[0030] Example 4 The difference between this embodiment and Embodiment 1 is that: In step (1), the weight ratio of Linglingxiang to Baizhi is 1:1.2; In step (2), the material-to-liquid ratio is 1:15 (g / mL), and the extraction is performed 3 times.

[0031] To verify that the above extract can be used to treat seborrheic dermatitis and has a non-toxic and harmless effect, the present invention conducted the following experiment on the finished dry powder in Example 1.

[0032] 1. Animal experiment toxicity verification of the aqueous extract of *Linglingxiang* and *Baizhi* for the treatment of seborrheic dermatitis: SPF-grade white male guinea pigs, 5 weeks old, weighing 350g-400g, were purchased from Shaanxi Junxing Biotechnology Co., Ltd., license number: SCXK (Shaanxi) 2022-01. They were randomly divided into 4 groups (n=4): control group, 2% ketoconazole group, high-dose group, and low-dose group. The extract powder from Example 1 was dissolved in pure water to prepare high- and low-dose drug concentrations of 10.0g / mL and 5.0g / mL, respectively.

[0033] One week after the guinea pigs were fed an adaptive diet, the hair on both sides of their backs was removed using an electric shaver and depilatory cream. Each shaved area was 2×3cm. 2 Afterwards, wipe the hair removal area with saline solution to confirm that the skin is free of damage, erythema, and edema, and then set it aside (if any skin abnormalities are found, remove the guinea pig). Administer the medication once daily for 14 consecutive days, applying it evenly to the hair removal area on the back, covering an area of ​​2cm². 3cm, dosage is 0.3g / cm 2 (Same dosage as in the treatment experiment).

[0034] ① Observation of local skin irritation: Administer the medication continuously for 14 days, allowing it to be absorbed naturally after administration without bandaging. Observe the skin condition daily. Based on the skin irritation scoring criteria in the "Cosmetic Safety Technical Specifications," scores were given from two dimensions: "erythema and eschar" and "edema," as shown in Table 1.

[0035] Table 1 The average stimulation score of each group of guinea pigs was recorded daily (total score of 6 guinea pigs in each group / 6); after the administration was completed, the "cumulative stimulation index" (total average stimulation score of each day / 14) was calculated over 14 days. If the cumulative stimulation index was ≤0.5, it was judged as "no stimulation"; 0.5 < index ≤2.0 was "mild stimulation"; and >2.0 was "moderate / severe stimulation".

[0036] Among them, the skin appearance results of 4 groups of guinea pigs after 14 days of drug treatment are as follows: Figure 1 As shown, through Figure 1 It can be seen that the test substance, administered topically to the skin at both high and low doses for 14 consecutive days, did not cause any irritation or damage to the guinea pig skin, and had no local toxic side effects. Its skin safety is comparable to that of 2% ketoconazole, a first-line positive control drug in clinical practice. It has a safe basis for long-term topical administration to the skin, providing core toxicological safety support for the local treatment of skin diseases such as seborrheic dermatitis.

[0037] The average stimulation scores of each group of guinea pigs are shown in Table 2. It can be seen that all four groups of guinea pigs were determined to be "non-irritating" after the administration of the drug.

[0038] Table 2: ② Observation of systemic toxicity and irritation: Record the food intake and weight changes of the guinea pigs. The food intake was recorded every two days based on the difference in the weight of the remaining feed in each group. The weight of the guinea pigs was weighed using an electronic balance, and the weight change rate of each group of mice was calculated as ((daily weight - initial weight) / initial weight × 100%). If the weight continued to decrease, it indicated that there might be systemic toxicity.

[0039] Changes in food intake and body weight in the four groups of guinea pigs are as follows: Figure 2 As shown, after 14 consecutive days of topical administration of the test substance to the skin, the guinea pigs' weight gain and feeding behavior were completely consistent with those of the blank control group and the positive drug control group, with no systemic toxicity related to the test substance, demonstrating excellent safety.

[0040] ③ Skin tissue pathological examination: On day 14 after drug administration, skin tissue from the dorsal area of ​​the guinea pig was taken, fixed in 10% paraformaldehyde, stained with hematoxylin and eosin (HE), and observed under a microscope. The experimental results are as follows: Figure 3 As shown, the test substance, administered topically to the skin at both high and low doses for 14 consecutive days, did not cause any pathological damage to the guinea pig skin tissue, nor did it cause any local irritation or tissue toxicity. Its skin safety is comparable to that of 2% ketoconazole, a first-line positive drug in clinical practice, and it has a safe basis for long-term topically administered skin treatment.

[0041] In summary, the extract powder of the present invention is safe and non-toxic for the local treatment of skin diseases such as seborrheic dermatitis.

[0042] To verify that the above extract can be used to treat seborrheic dermatitis, the present invention conducted the following experiment on the finished dry powder from Example 1.

[0043] 2. Efficacy verification of the aqueous extract of *Linglingxiang* and *Baizhi* in treating seborrheic dermatitis in animal experiments: SPF-grade white male guinea pigs, 5 weeks old, weighing 350g-400g, were purchased from Shaanxi Junxing Biotechnology Co., Ltd., license number: SCXK (Shaanxi) 2022-01. They were randomly divided into 5 groups (n=5): Control group, Model group, 2% ketoconazole group, High-dose group, and Low-dose group.

[0044] One week after adaptive feeding, a seborrheic dermatitis model was established. Hair was removed from both sides of the guinea pigs' backs using an electric shaver and depilatory cream, with each shaved area measuring 2×3 cm. 2 Inoculation was performed the day after shaving. Before inoculation, the skin on the back of the guinea pig was rubbed with a cotton swab until it became red but without bleeding. Except for the control group, the other four groups were treated with a modified 7-day bacterial-lipid topical application method to apply the prepared bacterial suspension once a day for 7 consecutive days.

[0045] The extract powder from Example 1 was dissolved in pure water to prepare high-concentration and low-concentration drug groups, at concentrations of 10.0 g / mL and 5.0 g / mL, respectively. After modeling, the corresponding drugs were applied to the skin lesions of the 2% ketoconazole group, the high-concentration group, and the low-concentration group, with an application area of ​​2 cm². Apply the ointment to a depth of 3cm twice daily for 7 consecutive days. During the experiment, take photographs and weigh the ointment every two days.

[0046] Observe the progress of modeling and skin lesions daily; visually observe and assess the degree of changes in erythema, papules and scales on the skin lesion area every day, and score them by clinical score (the sum of erythema, scale and papule scores) from 0 to 6.

[0047] On day 14, abscess tissue was collected from the back of the guinea pigs, fixed in 10% paraformaldehyde, stained with hematoxylin and eosin (HE) as usual, and observed under a microscope. Blood was collected from the heart to obtain guinea pig serum, and the expression levels of inflammatory factors in the serum were measured using enzyme-linked immunosorbent assay (ELISA).

[0048] Experimental results are as follows Figure 4-7 As shown.

[0049] pass Figure 4 It can be seen that the high-concentration group, low-concentration group, and 2% ketoconazole group all significantly improved seborrheic dermatitis. Compared with the model group, the skin lesion morphology scores of the treatment groups, such as erythema, scaling, and papules, were significantly reduced, and the skin lesion symptoms were significantly relieved. The high-concentration and low-concentration groups rapidly and significantly improved the core skin lesion symptoms such as erythema and scaling in the model guinea pigs, accelerating the recovery of the skin's normal physiological state. The high-concentration group showed significantly better onset of action, degree of skin lesion improvement, and skin recovery effect than the low-concentration group. Furthermore, the high-concentration group also had a faster onset of action than the 2% ketoconazole positive control group, and the degree of skin lesion recovery at 14 days of administration was significantly better than the positive control group, indicating that high-dose extracts have a potent and rapid therapeutic effect on seborrheic dermatitis.

[0050] Comparison chart of quantitative results of skin lesion scoring ( Figure 5 (B) shows dynamic changes in the overall appearance of the skin lesions that are completely consistent with previous findings. Through... Figure 5It can be seen that both the high-concentration and low-concentration drug groups have significant therapeutic effects on seborrheic dermatitis and can improve the skin lesion symptoms of model guinea pigs in a dose-dependent manner. The high-concentration drug group has a faster onset of action and better therapeutic effect, and its overall efficacy is higher than that of the first-line positive drug 2% ketoconazole in clinical practice.

[0051] pass Figure 6 It can be seen that the model group rats showed obvious pathological damage to their skin tissue, which was characterized by irregular thickening of the epidermis, excessive keratinization of the stratum corneum with focal desquamation, massive infiltration of inflammatory cells in the dermis, and inflammatory accumulation around the hair follicles, which are typical pathological features of seborrheic dermatitis. After the intervention of the extract, the thickening of the epidermis of the rats in the treatment group was significantly improved, the abnormal keratinization of the stratum corneum was significantly relieved, the degree of inflammatory cell infiltration in the dermis and around the hair follicles was greatly reduced, and the skin tissue structure tended to be intact and close to the normal skin morphology.

[0052] Figure 7 (E) represents the serum IL-1β expression level. IL-1β is a core early factor initiating inflammation in seborrheic dermatitis and is the "master switch" that triggers the local inflammatory cascade response in the skin. It can directly induce abnormal proliferation of keratinocytes and damage to the skin barrier, and is the core cause of erythema and scaling.

[0053] The blank control group showed that serum IL-1β remained stable at a normal physiological low level of approximately 25 pg / mL, with no abnormal inflammatory activation. The model group showed a significantly higher serum IL-1β level of approximately 45 pg / mL compared to the blank control group (P<0.001), confirming that the modeling procedure successfully activated the inflammatory pathway in the model animals, consistent with the pathological characteristics of seborrheic dermatitis. In contrast, the 2% ketoconazole positive control group showed a decrease in serum IL-1β level to approximately 28 pg / mL, a significantly lower level than the model group (P<0.001), returning to near-normal levels, confirming that the first-line clinical treatment drug can effectively inhibit the abnormal activation of inflammatory initiating factors. In the high-concentration group of the test drug, serum IL-1β level decreased to approximately 30 pg / mL, a significantly lower level than the model group (P<0.001), with an anti-inflammatory effect essentially equivalent to the positive drug, confirming that the test drug can fundamentally block the inflammatory initiation process of seborrheic dermatitis.

[0054] Figure 7 (F) represents the serum IL-6 expression level. IL-6 is a core inflammatory amplifying factor that connects innate immunity and adaptive immunity. It is also a key driver that induces the differentiation of pathogenic Th17 cells and is a core driving factor for the transformation of seborrheic dermatitis from acute inflammation to chronic protracted and recurrent seborrheic dermatitis.

[0055] The blank control group showed that serum IL-6 remained stable at a normal physiological low level of approximately 85 pg / mL. Meanwhile, the model group showed a highly significant increase in serum IL-6 levels to approximately 170 pg / mL compared to the blank control group (P<0.001), confirming that the model not only induced acute inflammation but also activated signaling pathways related to chronic inflammation, and the model conformed to the disease progression characteristics of seborrheic dermatitis. In the 2% ketoconazole positive control group, serum IL-6 levels decreased to approximately 100 pg / mL after administration, a highly significant downregulation compared to the model group (P<0.001). In contrast, the high-concentration test drug group showed a significantly downregulation of serum IL-6 levels to approximately 130 pg / mL after administration, a highly significant downregulation compared to the model group (P<0.001), confirming that the test drug can effectively inhibit the inflammatory amplification process and block upstream induction signals for Th17 cell differentiation, demonstrating the potential to improve chronic inflammation and reduce the risk of recurrence.

[0056] Figure 7 (G) represents the serum IL-17 expression level. IL-17 is mainly secreted by differentiated and mature Th17 cells. It is a core effector factor in the chronicity of seborrheic dermatitis and irreversible damage to the skin barrier. It can directly inhibit the synthesis of core structural proteins of the skin barrier such as filaggrin and naegrin, while maintaining the continuous activation of the inflammatory cascade response. It is the core molecular basis for the recurrence of the disease.

[0057] The blank control group showed that serum IL-17 was stably maintained at a normal physiological low level of about 32 pg / mL. The serum IL-17 level in the model group was significantly higher than that in the blank control group, reaching approximately 48 pg / mL (P<0.001), confirming that the model successfully induced abnormal activation of the Th17 pathway and constructed a stable chronic seborrheic dermatitis model. In the 2% ketoconazole positive control group, the serum IL-17 level decreased to approximately 35 pg / mL after administration, which was significantly downregulated compared to the model group (P<0.001), returning to near-normal levels. In the high-concentration group of the test drug, the serum IL-17 level decreased to approximately 37 pg / mL after administration, which was significantly downregulated compared to the model group (P<0.001), confirming that the test drug can effectively block the core effector of chronic inflammation, relieve the inhibition of skin barrier synthesis by inflammatory factors, and create conditions for barrier repair.

[0058] Figure 7 (H) represents the serum TNF-α expression level. TNF-α is the core amplifying factor in the inflammatory cascade of seborrheic dermatitis. It can directly induce skin vasodilation, erythema, edema, and itching. At the same time, it stimulates excessive secretion of sebaceous glands, providing nutrition for the proliferation of Malassezia, forming a vicious cycle of "inflammation → increased sebum → microbial proliferation → aggravated inflammation". It is the direct cause of core symptoms such as erythema, itching, and oily scaling.

[0059] The blank control group showed that serum TNF-α was stably maintained at a normal physiological low level of approximately 150 pg / mL. The serum TNF-α level in the model group was significantly increased to approximately 230 pg / mL compared with the blank control group (P<0.001), further confirming that the inflammatory activation state of the model was highly matched with the clinicopathological features of seborrheic dermatitis. In the 2% ketoconazole positive control group, the serum TNF-α level decreased to approximately 155 pg / mL after administration, which was significantly downregulated compared with the model group (P<0.001), and completely recovered to the normal physiological level. In contrast, the high concentration group of the test drug decreased the serum TNF-α level to approximately 170 pg / mL after administration, which was significantly downregulated compared with the model group (P<0.001), confirming that the test drug can effectively block the vicious cycle of inflammation amplification and simultaneously improve core clinical symptoms such as erythema, itching, and greasy scaling.

[0060] Based on the above results, the aqueous extracts of *Eupatorium fortunei* and *Angelica dahurica* of the present invention can exert synergistic anti-inflammatory effects through multiple targets, and the specific mechanism of action is as follows: (1) Inhibit the release of inflammatory mediators: The aqueous extract of Linglingxiang-Angelica dahurica can inhibit the expression of inflammatory factors such as TNF-α, il-1β, il-6, and il-17, reduce the accumulation of inflammatory mediators at the site of skin inflammation, and thus alleviate inflammatory symptoms such as skin erythema, swelling, and itching.

[0061] (2) Blocking inflammatory signaling pathways: By inhibiting the expression of upstream pro-differentiation cytokines (especially IL-6 and IL-1β), targeting and inhibiting the IL-6 / JAK-STAT3 axis, and inhibiting the synthesis and secretion of downstream pro-inflammatory factors such as IL-17, abnormal differentiation and inflammatory response of TH17 cells are inhibited.

[0062] (3) Regulating skin barrier function: The active ingredients in the aqueous extract of Angelica dahurica can reduce the excessive proliferation and differentiation of stratum corneum cells, repair the damaged skin barrier, reduce the damage to the skin caused by external stimuli, and inhibit the excessive secretion of sebaceous glands, improve the oily skin lesions of seborrheic dermatitis, fundamentally reduce the inducing factors of inflammatory response, improve the treatment effect, and reduce the recurrence rate. This effect makes up for the shortcomings of existing drugs that only focus on anti-inflammation and neglect skin barrier repair, and further enhances the clinical application value.

[0063] In summary, the present invention has the following effects: 1. It can significantly downregulate the abnormal expression of four core pro-inflammatory factors: IL-1β, IL-6, IL-17, and TNF-α, covering the entire chain of seborrheic dermatitis: "inflammation initiation → inflammation amplification → chronic effects → vicious cycle maintenance." This confirms its multi-target synergistic anti-inflammatory mechanism, rather than single-pathway intervention, and its anti-inflammatory effect is comparable to that of 2% ketoconazole, a first-line clinical positive control drug. Furthermore, the extract of this invention not only inhibits acute inflammatory factors (IL-1β, TNF-α) related to innate immunity, rapidly improving skin lesion symptoms, but also blocks the chronic inflammatory pathway (IL-6, IL-17) related to Th17 differentiation, fundamentally addressing the industry pain point of chronic and recurrent seborrheic dermatitis, highly matching the clinical treatment needs of seborrheic dermatitis.

[0064] 2. This invention utilizes the scientific combination of Linglingxiang and Baizhi, along with an optimized water extraction process, to fully preserve the synergistic effects of their water-soluble anti-inflammatory and skin barrier repair active ingredients. By relying on the synergistic effect of the combination, it effectively inhibits the inflammatory response of seborrheic dermatitis, thereby alleviating typical symptoms such as skin erythema, itching, and desquamation, and achieving the repair and improvement of skin lesions.

[0065] 3. This approach reduces the potential side effects of single-component drugs, improves medication safety and patient tolerability, reduces disease recurrence, overcomes the limitations of existing treatment options, and provides a safe, effective, low-cost, and easily promoted anti-inflammatory treatment product and related technical solutions for seborrheic dermatitis. Simultaneously, through the rational combination of *Eupatorium fortunei* and *Angelica dahurica*, the flavonoid glycosides in *Eupatorium fortunei* can effectively alleviate the photosensitivity toxicity caused by components in *Angelica dahurica*.

[0066] 4. The water extraction process can retain water-soluble active ingredients to the greatest extent, resulting in good biocompatibility, excellent skin absorption, and strong tolerability. This invention employs an optimized water extraction process that is simple and easy to operate, requiring no complex equipment or expensive reagents. Multiple water extractions improve raw material utilization and reduce production costs. This extraction method can maximize the retention of active ingredients, ensuring stable therapeutic effects. The resulting water extract powder is easy to store, transport, and process into formulations, and can be flexibly prepared into various dosage forms such as topical creams, lotions, gels, and oral capsules and granules to meet the needs of different patients. It is also easy for large-scale industrial production and clinical application.

[0067] 5. This invention precisely targets the pathogenesis of seborrheic dermatitis, combining the pharmacological advantages of Eupatorium fortunei and Angelica dahurica. For the first time, the two are combined to prepare an aqueous extract and applied to the anti-inflammatory treatment of seborrheic dermatitis, filling the gap in the existing technology that does not involve this combination application. Compared with existing single plant extracts, it solves the problems of limited anti-inflammatory effect and limited application scenarios. Compared with existing chemical drugs, it solves the defects of large side effects, easy recurrence and poor patient tolerance, while taking into account efficacy, safety and practicality.

[0068] 6. Using Linglingxiang and Baizhi that meet the pharmacopoeia standards as raw materials, the core weight ratio is 1:1-2 (1:1 is optimal). This ratio achieves synergistic effects of the active ingredients of the two, taking into account both anti-inflammatory effects and safety, and alleviating the photosensitivity toxicity of Baizhi alone.

[0069] 7. Repairs the skin barrier, reduces excessive keratinization of skin tissue, and provides targeted treatment for seborrheic dermatitis and dandruff.

[0070] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

Claims

1. A method for preparing extracts of *Eupatorium fortunei* and *Angelica dahurica*, characterized in that, Includes the following steps: (1) Weighing raw materials Weigh out Linglingxiang and Baizhi according to a weight ratio of 1:1-2, and set aside. (2) Extraction by decoction The raw materials in step (1) are mixed with water at a ratio of 1:10-15 (g / mL) and heated, and then extracted by decoction 2-3 times. (3) Filtration After each extraction in step (2), perform solid-liquid separation, collect the filtrates from each extraction, and mix them for later use. (4) Finished product preparation The mixed filtrate from step (3) is concentrated and dried to obtain the finished dry powder.

2. The preparation method according to claim 1, characterized in that, In step (1), Linglingxiang and Baizhi are cleaned, dried, and pulverized before being weighed.

3. The preparation method according to claim 1, characterized in that, In step (2), the material-to-liquid ratio is 1:10 (g / mL), and extraction is performed twice.

4. The preparation method according to claim 1 or 3, characterized in that, In step (2), after heating to boiling, maintain a gentle boil and stir every 5-10 minutes; Calculate the extraction time after boiling, and the extraction time is 30-50 minutes each time.

5. The preparation method according to claim 1, characterized in that, In step (3), the solid and liquid are filtered by vacuum filtration.

6. The preparation method according to claim 1, characterized in that, In step (4), the mixed filtrate is concentrated to a relative density of 1.10-1.20 (25°C) and then dried.

7. An extract, characterized in that, It is prepared by any of the preparation methods in claims 1-6.

8. The use of the extract according to claim 7 in the preparation of pharmaceuticals and topical products for treating inflammatory skin diseases and repairing the skin barrier.

9. The application according to claim 8, characterized in that, Inflammatory skin diseases include seborrheic dermatitis.

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