Application of hesperidin derivative in preparation of product for treating skin lesion type skin diseases

Abstract

The invention relates to the technical field of medicines and daily chemical products, in particular to application of a hesperidin derivative in preparation of a product for treating skin lesion skin diseases. The hesperidin derivative is selected from glucosyl hesperidin, methyl hesperidin chalcone or neohesperidin dihydrochalcone. The product has a remarkable effect on treating psoriasis, atopic dermatitis, irritant dermatitis and sensitive skin and repairing scars, can be prepared into various external dosage forms, is small in side effect and high in safety, is extracted from natural plants, can reduce hormone dependence, and provides a safer and more effective solution for treating skin lesion skin diseases.

Description

Technical Field

[0001] The present invention relates to the technical fields of medicine and daily chemical products, and particularly to the application of hesperidin derivatives in the preparation of products for treating skin-lesion dermatoses. Background Art

[0002] Skin-lesion dermatoses is a general term for a large category of skin diseases, mainly referring to various forms of skin damage. Skin damage (skin lesions) is an important basis for diagnosing skin diseases, and these damages can be manifested as primary skin lesions and secondary skin lesions.

[0003] Common skin-lesion dermatoses include psoriasis, dermatitis, eczema, skin lesions caused by sensitive skin or scars, etc. Currently commonly used topical medications such as glucocorticoids, antibiotics, antihistamines, etc., although can relieve symptoms to a certain extent, may produce drug resistance, dependence, and adverse reactions such as skin atrophy and pigmentation after long-term use. Moreover, for some severe skin-lesion dermatoses, the efficacy of topical medications is often limited and it is difficult to achieve a radical cure. Oral medications such as immunosuppressants, biological agents, etc., although have a certain effect on controlling the condition, have systemic side effects, such as liver and kidney function damage, increased risk of infection caused by immunosuppression, etc., which limit their long-term use.

[0004] In view of the many problems existing in the current treatment and nursing means, there is an urgent need to develop a new treatment method, drug or nursing product that can more effectively treat skin-lesion dermatoses, relieve symptoms, reduce the recurrence rate, and at the same time reduce adverse reactions and side effects. Summary of the Invention

[0005] To solve or partially solve the problems existing in the related technologies, the present invention provides an application of hesperidin derivatives in the preparation of products for treating skin-lesion dermatoses.

[0006] The present invention provides an application of hesperidin derivatives in the preparation of products for treating skin-lesion dermatoses, and the hesperidin derivatives are selected from glucosyl hesperidin, methyl hesperidin chalcone or neohesperidin dihydrochalcone.

[0007] Further, the concentration of the hesperidin derivative in the product is 0.01% - 5%.

[0008] Further, the skin-lesion dermatosis is psoriasis.

[0009] Further, the concentration of the hesperidin derivative in the product is 0.1% - 2%.

[0010] Further, the skin-lesion dermatoses are atopic dermatitis, irritant dermatitis, skin damage caused by sensitive skin or scars.

[0011] Furthermore, the concentration of hesperidin derivatives in the product is 0.1% - 3%.

[0012] Furthermore, the product is the hesperidin derivative, or the product comprises: the hesperidin derivative and a pharmaceutically or cosmetically acceptable carrier.

[0013] Furthermore, the dosage form of the product includes: ointment, spray, liniment, patch, emulsion, gel or lotion.

[0014] Furthermore, the product exerts a therapeutic effect through at least one of the following mechanisms: Regulating the expression levels of S100A8, S100A9, and K6 mRNA; Reducing the expression levels of IL-1α, IL-1β, CXCL1, and CXCL2; Reducing the expression levels of IL-17A, IL-22, and IL-23A; Reducing the expression levels of VEGF-A and ICAM-1.

[0015] Furthermore, the product further contains at least one active ingredient selected from glucocorticoids, calcineurin inhibitors, antibiotics, antihistamines, and immunosuppressants. The application of the hesperidin derivative provided by the present invention in the preparation of a product for treating skin lesion dermatoses may include the following beneficial effects: 1. The product can significantly inhibit psoriasis-related inflammatory factors (IL-17A, IL-23A, S100A8 / A9), reduce epidermal thickening and scale formation, and has a good improvement effect on psoriasis. The product effectively relieves skin erythema, edema, and scratching behavior by reducing Th2-type cytokines (IL-4, IL-13) and itch-related chemokines (CXCL1 / CXCL2), and has a good therapeutic effect on atopic dermatitis. The efficacy is comparable to that of glucocorticoids but with higher safety. The product has a good therapeutic effect on irritant dermatitis by inhibiting TPA-induced ear inflammation and edema and reducing the expression of pro-inflammatory factors such as IL-1β and VEGF-A. The product can significantly reduce the TEWL value, restore the skin barrier function, reduce abnormal thickening of the stratum corneum and inflammatory reactions, and can also inhibit the excessive proliferation of fibroblasts, reduce scar tissue formation, and promote wound healing, and has a good therapeutic effect on sensitive skin and scar repair.

[0016] 2. The product can be made into various external dosage forms such as ointments, gels, emulsions, sprays, patches, etc., which is convenient for local precise drug delivery and improves patient compliance. Compared with traditional hormonal drugs (such as glucocorticoids and calcineurin inhibitors), hesperidin derivatives can exert significant efficacy within the concentration range of 0.1% - 3%, avoiding adverse reactions such as hormone-dependent dermatitis and skin atrophy. It can partially replace hormonal drugs, reduce patients' dependence on glucocorticoids, and is especially suitable for the long-term management of chronic skin diseases (such as atopic dermatitis and psoriasis).

[0017] 3. Hesperidin derivatives are derived from plant extracts, with the characteristics of natural safety and high biocompatibility, and are suitable for long-term use.

[0018] The product provided by this patent has significant effects in the treatment of psoriasis, atopic dermatitis, irritant dermatitis, sensitive skin, and scar repair. It can be made into various external dosage forms, with few side effects and high safety. Moreover, it is derived from natural plant extracts, which can reduce hormone dependence and provide a safer and more effective solution for the treatment of skin lesion dermatoses. Brief Description of the Drawings

[0019] Figure 1 It is a bar graph showing the influence of compound A at different concentrations on the skin PASI score in psoriasis mice; compared with the IMQ + matrix group, *P < 0.05, **P < 0.01, ***P < 0.001; compared with the IMQ + calcipotriol group, ## P < 0.01.

[0020] Figure 2 It is a physical picture showing the changes in the dorsal skin of psoriasis mice with compound A at different concentrations.

[0021] Figure 3 It is a HE staining picture of histopathological observation of skin lesions in psoriasis mice with compound A at different concentrations.

[0022] Figure 4 It is a bar graph showing the relative mRNA levels of S100A8, S100A9, K6, and K16 in the skin of psoriasis mice with compound A at different concentrations; compared with the IMQ + matrix group, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the IMQ + calcipotriol group, ## P < 0.01, ### P < 0.001, #### P < 0.0001.

[0023] Figure 5Bar graph of the relative mRNA levels of IL-17A, IL-22, and IL-23A in the skin of psoriasis mice at different concentrations of Compound A; compared with the IMQ + vehicle group, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the IMQ + calcipotriol group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0024] Figure 6 Bar graph of the relative mRNA levels of IL-1α, IL-1β, CXCL1, and CXCL2 in the skin of psoriasis mice at different concentrations of Compound A; compared with the IMQ + vehicle group, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the IMQ + calcipotriol group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0025] Figure 7 Bar graph of the relative mRNA levels of VEGF-A and ICAM-1 in the skin of psoriasis mice at different concentrations of Compound A; compared with the IMQ + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001; compared with the IMQ + calcipotriol group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0026] Figure 8 Bar graph of the effect of different concentrations of Compound B on the PASI score of the skin in psoriasis mice; compared with the IMQ + vehicle group, *P < 0.05, **P < 0.01.

[0027] Figure 9 Photograph of the dorsal skin changes in psoriasis mice at different concentrations of Compound B.

[0028] Figure 10 HE staining images of the histopathological observation of the skin lesions in psoriasis mice at different concentrations of Compound B.

[0029] Figure 11 Bar graph of the relative mRNA levels of S100A8, S100A9, K6, and K16 in the skin of psoriasis mice at different concentrations of Compound B; compared with the IMQ + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001; compared with the IMQ + calcipotriol group, # P < 0.05, ##P < 0.01, ### P < 0.001.

[0030] Figure 12 is a bar graph of the relative mRNA levels of IL-17A, IL-22, and IL-23A in the skin of psoriasis mice at different concentrations of compound B; compared with the IMQ + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001; compared with the IMQ + calcipotriol group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0031] Figure 13 is a bar graph of the relative mRNA levels of IL-1α, IL-1β, CXCL1, and CXCL2 in the skin of psoriasis mice at different concentrations of compound B; compared with the IMQ + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001; compared with the IMQ + calcipotriol group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0032] Figure 14 is a bar graph of the relative mRNA levels of VEGF-A and ICAM-1 in the skin of psoriasis mice at different concentrations of compound B; compared with the IMQ + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001; compared with the IMQ + calcipotriol group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0033] Figure 15 is a bar graph of the effect of different concentrations of compound C on the PASI score of the skin in psoriasis mice; compared with the IMQ + vehicle group, *P < 0.05, **P < 0.01.

[0034] Figure 16 is a physical picture of the changes in the dorsal skin of psoriasis mice at different concentrations of compound C.

[0035] Figure 17 is a HE staining picture of the histopathological observation of the skin lesions in psoriasis mice at different concentrations of compound C.

[0036] Figure 18Bar graph of the relative mRNA levels of S100A8, S100A9, K6, and K16 in the skin of psoriatic mice treated with different concentrations of Compound C; compared with the IMQ + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001; compared with the IMQ + calcipotriol group, # P < 0.05, ## P < 0.01, ### P < 0.001. Figure 19 Bar graph of the relative mRNA levels of IL-17A, IL-22, and IL-23A in the skin of psoriatic mice treated with different concentrations of Compound C; compared with the IMQ + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001; compared with the IMQ + calcipotriol group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0037] Figure 20 Bar graph of the relative mRNA levels of IL-1α, IL-1β, CXCL1, and CXCL2 in the skin of psoriatic mice treated with different concentrations of Compound C; compared with the IMQ + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001; compared with the IMQ + calcipotriol group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0038] Figure 21 Bar graph of the relative mRNA levels of VEGF-A and ICAM-1 in the skin of psoriatic mice treated with different concentrations of Compound C; compared with the IMQ + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001; compared with the IMQ + calcipotriol group, # P < 0.05, ## P < 0.01, ### P < 0.001.

[0039] Figure 22 Bar graph of the effects of different concentrations of Compound A on the dermatitis severity score and scratching frequency in the skin of atopic dermatitis mice; compared with the DNFB + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the DNFB + eloson group, # P < 0.05.

[0040] Figure 23Bar graph of the relative mRNA levels of S100A8, S100A9, and K6 in the skin of atopic dermatitis mice treated with different concentrations of compound A; compared with the DNFB + vehicle group, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the DNFB + eloson group, ## P < 0.01, ### P < 0.001.

[0041] Figure 24 Bar graph of the relative mRNA levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in the skin of atopic dermatitis mice treated with different concentrations of compound A; compared with the DNFB + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the DNFB + eloson group, # P < 0.05, ## P < 0.01.

[0042] Figure 25 Bar graph of the relative mRNA levels of IL-17A, IL-22, and IL-23A in the skin of atopic dermatitis mice treated with different concentrations of compound A; compared with the DNFB + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the DNFB + eloson group, # P < 0.05, ## P < 0.01.

[0043] Figure 26 Bar graph of the relative mRNA levels of VEGF-A and ICAM-1 in the skin of atopic dermatitis mice treated with different concentrations of compound A; compared with the DNFB + vehicle group, **P < 0.01, ***P < 0.001; compared with the DNFB + eloson group, # P < 0.05.

[0044] Figure 27 Bar graph of the effects of different concentrations of compound B on the dermatitis severity score and scratching frequency in the skin of atopic dermatitis mice; compared with the DNFB + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0045] Figure 28 Bar graph of the relative mRNA levels of S100A8, S100A9, and K6 in the skin of atopic dermatitis mice treated with different concentrations of compound B; compared with the DNFB + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0046] Figure 29 Bar chart of the relative mRNA levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in the skin of atopic dermatitis mice with different concentrations of compound B; compared with the DNFB + vehicle group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0047] Figure 30 Bar chart of the relative mRNA levels of IL-17A, IL-22, and IL-23A in the skin of atopic dermatitis mice with different concentrations of compound B; compared with the DNFB + vehicle group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0048] Figure 31 Bar chart of the relative mRNA levels of VEGF-A and ICAM-1 in the skin of atopic dermatitis mice with different concentrations of compound B; compared with the DNFB + vehicle group, *P<0.05, **P<0.01, ***P<0.001.

[0049] Figure 32 Bar chart of the effects of different concentrations of compound C on the dermatitis severity score and scratching times in the skin of atopic dermatitis mice; compared with the DNFB + vehicle group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0050] Figure 33 Bar chart of the relative mRNA levels of S100A8, S100A9, and K6 in the skin of atopic dermatitis mice with different concentrations of compound C; compared with the DNFB + vehicle group, *P<0.05, **P<0.01, ***P<0.001.

[0051] Figure 34 Bar chart of the relative mRNA levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in the skin of atopic dermatitis mice with different concentrations of compound C; compared with the DNFB + vehicle group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0052] Figure 35 Bar chart of the relative mRNA levels of IL-17A, IL-22, and IL-23A in the skin of atopic dermatitis mice with different concentrations of compound C; compared with the DNFB + vehicle group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0053] Figure 36 Bar chart of relative mRNA levels of VEGF-A and ICAM-1 in the skin of atopic dermatitis mice with different concentrations of Compound C; compared with the DNFB + matrix group, *P<0.05, **P<0.01, ***P<0.001.

[0054] Figure 37 Bar chart of the effect of different concentrations of Compound A on the TEWL value of the skin of sensitive-skin mice; compared with the Tape + matrix group, **P<0.01, ***P<0.001; compared with the Tape + calcipotriol group, # P<0.05.

[0055] Figure 38 Bar chart of relative mRNA levels of S100A8, S100A9, K6 and K16 in the skin of sensitive-skin mice with different concentrations of Compound A; compared with the Tape + matrix group, ***P<0.001; compared with the Tape + calcipotriol group, # P<0.05.

[0056] Figure 39 Bar chart of relative mRNA levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1 and CXCL2 in the skin of sensitive-skin mice with different concentrations of Compound A; compared with the Tape + matrix group, **P<0.01, ***P<0.001, ****P<0.0001; compared with the Tape + calcipotriol group, # P<0.05, ## P<0.01.

[0057] Figure 40 Bar chart of relative mRNA levels of IL-17A, IL-22 and IL-23A in the skin of sensitive-skin mice with different concentrations of Compound A; compared with the Tape + matrix group, **P<0.01, ***P<0.001; compared with the Tape + calcipotriol group, # P<0.05, ## P<0.01.

[0058] Figure 41 Bar chart of relative mRNA levels of VEGF-A and ICAM-1 in the skin of sensitive-skin mice with different concentrations of Compound A; compared with the Tape + matrix group, **P<0.01, ***P<0.001; compared with the Tape + calcipotriol group, # P<0.05.

[0059] Figure 42Bar graph showing the effect of different concentrations of Compound B on the TEWL value of the skin of mice with sensitive skin; compared with the Tape + matrix group, *P < 0.05, **P < 0.01, ***P < 0.001.

[0060] Figure 43 Bar graph showing the relative mRNA levels of S100A8, S100A9, K6, and K16 in the skin of mice with sensitive skin at different concentrations of Compound B; compared with the Tape + matrix group, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0061] Figure 44 Bar graph showing the relative mRNA levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in the skin of mice with sensitive skin at different concentrations of Compound B; compared with the Tape + matrix group, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0062] Figure 45 Bar graph showing the relative mRNA levels of IL-17A, IL-22, and IL-23A in the skin of mice with sensitive skin at different concentrations of Compound B; compared with the Tape + matrix group, **P < 0.01, ***P < 0.001.

[0063] Figure 46 Bar graph showing the relative mRNA levels of VEGF-A and ICAM-1 in the skin of mice with sensitive skin at different concentrations of Compound B; compared with the Tape + matrix group, **P < 0.01, ***P < 0.001.

[0064] Figure 47 Bar graph showing the effect of different concentrations of Compound C on the TEWL value of the skin of mice with sensitive skin; compared with the Tape + matrix group, *P < 0.05, **P < 0.01, ***P < 0.001.

[0065] Figure 48 Bar graph showing the relative mRNA levels of S100A8, S100A9, K6, and K16 in the skin of mice with sensitive skin at different concentrations of Compound C; compared with the Tape + matrix group, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0066] Figure 49 Bar graph showing the relative mRNA levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in the skin of mice with sensitive skin at different concentrations of Compound C; compared with the Tape + matrix group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0067] Figure 50 Bar chart of the relative mRNA levels of IL-17A, IL-22, and IL-23A in the skin of mice with sensitive skin at different concentrations of Compound C; compared with the Tape + matrix group, *P < 0.05, **P < 0.01, ***P < 0.001.

[0068] Figure 51 Bar chart of the relative mRNA levels of VEGF-A and ICAM-1 in the skin of mice with sensitive skin at different concentrations of Compound C; compared with the Tape + matrix group, *P < 0.05, **P < 0.01, ***P < 0.001.

[0069] Figure 52 Bar chart of the changes in ear margin thickness of mice with irritant dermatitis at different concentrations of Compound A; on day 4, compared with the TPA + matrix group, **P < 0.01, ***P < 0.001; compared with the TPA + positive drug group, # P < 0.05.

[0070] Figure 53 Bar chart of the relative mRNA levels of S100A8, S100A9, and K6 in the skin of mice with irritant dermatitis at different concentrations of Compound A; compared with the TPA + matrix group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the TPA + positive drug group, # P < 0.05, ## P < 0.01.

[0071] Figure 54 Bar chart of the relative mRNA levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in the skin of mice with irritant dermatitis at different concentrations of Compound A; compared with the TPA + matrix group, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the TPA + positive drug group, # P < 0.05, ## P < 0.01.

[0072] Figure 55 Bar chart of the relative mRNA levels of IL-17A, IL-22, and IL-23A in the skin of mice with irritant dermatitis at different concentrations of Compound A; compared with the TPA + matrix group, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the TPA + positive drug group, # P < 0.05, ## P < 0.01.

[0073] Figure 56 Bar chart of the relative mRNA levels of VEGF-A and ICAM-1 in the skin of mice with irritant dermatitis at different concentrations of Compound A; compared with the TPA + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001; compared with the TPA + positive drug group, ## P < 0.01.

[0074] Figure 57 Bar chart of the changes in ear margin thickness in mice with irritant dermatitis at different concentrations of Compound B; on day 4, compared with the TPA + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001.

[0075] Figure 58 Bar chart of the relative mRNA levels of S100A8, S100A9, and K6 in the skin of mice with irritant dermatitis at different concentrations of Compound B; compared with the TPA + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001.

[0076] Figure 59 Bar chart of the relative mRNA levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in the skin of mice with irritant dermatitis at different concentrations of Compound B; compared with the TPA + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Compared with the TPA + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0077] Figure 60 Bar chart of the relative mRNA levels of IL-17A, IL-22, and IL-23A in the skin of mice with irritant dermatitis at different concentrations of Compound B; compared with the TPA + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0078] Figure 61 Bar chart of the relative mRNA levels of VEGF-A and ICAM-1 in the skin of mice with irritant dermatitis at different concentrations of Compound B; compared with the TPA + vehicle group, *P < 0.05, **P < 0.01.

[0079] Figure 62 Bar chart of the changes in ear margin thickness in mice with irritant dermatitis at different concentrations of Compound C; on day 4, compared with the TPA + vehicle group, *P < 0.05, **P < 0.01, ***P < 0.001.

[0080] Figure 63It is a bar graph of the relative mRNA levels of S100A8, S100A9, and K6 in the skin of mice with irritant dermatitis for the compound C at the same concentration; compared with the TPA + matrix group, *P < 0.05, **P < 0.01, ***P < 0.001.

[0081] Figure 64 It is a bar graph of the relative mRNA levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in the skin of mice with irritant dermatitis for the compound C at different concentrations; compared with the TPA + matrix group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0082] Figure 65 It is a bar graph of the relative mRNA levels of IL-17A, IL-22, and IL-23A in the skin of mice with irritant dermatitis for the compound C at different concentrations; compared with the TPA + matrix group, *P < 0.05, **P < 0.01, ***P < 0.001.

[0083] Figure 66 It is a bar graph of the relative mRNA levels of VEGF-A and ICAM-1 in the skin of mice with irritant dermatitis for the compound C at different concentrations. Compared with the TPA + matrix group, *P < 0.05, **P < 0.01. Detailed implementation mode

[0084] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0085] The embodiments of the present invention provide an application of hesperidin derivatives in the preparation of products for treating skin lesion dermatoses, and the hesperidin derivatives are selected from glucosyl hesperidin, methyl hesperidin chalcone, or neohesperidin dihydrochalcone.

[0086] The above-mentioned product for treating skin-lesion dermatosis may be the above-mentioned hesperidin derivatives, i.e., they are compounds, specifically glucosyl hesperidin, methyl hesperidin chalcone or neohesperidin dihydrochalcone. Alternatively, the product is a composition; the concentration of the hesperidin derivative in the product is 0.01% - 5%. Preferably, the concentration of the hesperidin derivative in the product is 0.1% - 3%. Specifically, the concentration of the hesperidin derivative in the product may be 0.1%, 0.25%, 0.5%, 1%, 2% or 3%. All the concentrations described in this application refer to mass concentrations. For the case where the product is a composition, the product includes: the above-mentioned hesperidin derivative, and a pharmaceutically or cosmetically acceptable carrier. The carrier refers to the components other than the active molecule in a pharmaceutical administration preparation or a cosmetical preparation, including excipients such as excipients, buffers, absorption promoters, emulsifiers, thickeners, surfactants, antioxidants, preservatives and flavors, etc. Specifically, such as glycerol, polydimethylsiloxane, caprylic / capric triglyceride, shea butter, cetearyl alcohol, etc. The dosage form of the product may include: ointment, spray, liniment, patch, emulsion, gel or lotion.

[0087] The above-mentioned skin-lesion dermatosis may be psoriasis. Further, the concentration of the hesperidin derivative in the above-mentioned product for treating skin-lesion dermatosis is 0.1% - 2%, and the specific concentration may be 0.1%, 0.25%, 0.5% or 2%. The concentration of the hesperidin derivative is more preferably 0.5% - 2%, and most preferably 0.5%.

[0088] The above-mentioned skin-lesion dermatosis is atopic dermatitis, irritant dermatitis, skin damage caused by sensitive skin or scars. Further, the concentration of the hesperidin derivative in the above-mentioned product for treating skin-lesion dermatosis is 0.1% - 3%, and the specific concentration may be 0.1%, 0.25%, 0.5%, 1% or 3%. The concentration of the hesperidin derivative is more preferably 0.5% - 1%, and most preferably 0.5%.

[0089] The above-mentioned product for treating skin-lesion dermatosis may exert a therapeutic effect through at least one of the following mechanisms: Regulating the expression levels of S100A8, S100A9, and K6 mRNA; Reducing the expression levels of IL-1α, IL-1β, CXCL1, and CXCL2; Reducing the expression levels of IL-17A, IL-22, and IL-23A; Reducing the expression levels of VEGF-A and ICAM-1.

[0090] Preferably, the above-mentioned product for treating skin-lesion dermatosis further contains at least one active ingredient selected from glucocorticoids, calcineurin inhibitors, antibiotics, antihistamines, and immunosuppressants. The technical solution of the present invention will be further described below in conjunction with specific embodiments: I. The sources of raw materials involved in the following examples are as follows: Glucosyl hesperidin: Scentiva (Guangzhou) Trading Co., Ltd., 100 g / bag Methyl hesperidin chalcone: Shanghai Aladdin Biochemical Technology Co., Ltd., 100 g / bag Neohesperidin dihydrochalcone: Xi'an Luteng Biotechnology Co., Ltd., 100 g / bag Calcipotriol ointment: LEO Laboratories Limited 5% Imiquimod: Sichuan Mingxin Pharmaceutical Co., Ltd., 3 g / branch 2,4-Dinitrofluorobenzene DNFB: purity ≥ 98% (GC), from Shanghai Aladdin Reagent Co., Ltd., product number F475701-10g Eloson cream: Bayer Healthcare Shanghai Co., Ltd. Bakelite silicone gel: produced by Advanced Biotechnology Co., Ltd., 15 g / branch II. The matrices in the following examples are all configured according to the following method: Matrix composition: Glycerol (4%), polydimethylsiloxane (1.5%), caprylic / capric triglyceride (1.6%), shea butter (BUTYROSPERMUM PARKII) fruit butter (1.6%), glyceryl stearate (1.3%), cetearyl alcohol (0.5%), 1,2-hexanediol (0.43%), octanoyl hydroxamic acid (0.055%), ethylhexylglycerin (0.055%), carbomer (0.08%), and the balance of water.

[0091] Configuration method: 1. Put polydimethylsiloxane, caprylic / capric triglyceride, shea butter (BUTYROSPERMUM PARKII) fruit butter, glyceryl stearate, and cetearyl alcohol into a beaker, heat to 75 - 80 °C until completely melted, and stir evenly.

[0092] 2. Put water, glycerol, and carbomer into a beaker, heat to 80 - 85 °C, keep warm for 15 - 20 min, and then homogenize evenly.

[0093] 3. Pour the oil phase in step 1 into the water phase beaker in step 2, and homogenize until evenly mixed.

[0094] 4. Stir and cool to below 45 °C, add 1,2-hexanediol, octanoyl hydroxamic acid, and ethylhexylglycerin, and stir evenly.

[0095] III. Configuration method of hesperidin derivatives with different concentrations Calculate the weight ratio of hesperidin derivatives and matrix according to the preset concentration, add hesperidin derivatives to the matrix while stirring, and homogenize for 3 - 5 min until the hesperidin derivatives are evenly dispersed and dissolved in the matrix. Configure according to this method: Glucohesperidin with concentrations of 0.01 wt%, 0.1 wt%, 0.25 wt%, 0.5 wt%, 2 wt%, 3 wt% and 5 wt% are respectively denoted as: 0.01% Compound A, 0.1% Compound A, 0.25% Compound A, 0.5% Compound A, 2% Compound A, 3% Compound A and 5% Compound A.

[0096] Neohesperidin dihydrochalcone with concentrations of 0.01 wt%, 0.1 wt%, 0.25 wt%, 0.5 wt%, 2 wt%, 3 wt% and 5 wt% are respectively denoted as: 0.01% Compound B, 0.1% Compound B, 0.25% Compound B, 0.5% Compound B, 2% Compound B, 3% Compound B and 5% Compound B.

[0097] Methyl hesperidin chalcone with concentrations of 0.01 wt%, 0.1 wt%, 0.25 wt%, 0.5 wt%, 2 wt%, 3 wt% and 5 wt% are respectively denoted as: 0.01% Compound C, 0.1% Compound C, 0.25% Compound C, 0.5% Compound C, 2% Compound C, 3% Compound C and 5% Compound C.

[0098] Example 1 Experiment on the Improvement of Psoriasis Mouse Model by Hesperidin Derivatives 1. Experimental Materials 1.1 Experimental Animals Healthy female, SPF - level clean C57 mice, 6 - 8 weeks old.

[0099] 1.2 Experimental Reagents 5% Imiquimod (IMQ); matrix; calcipotriol ointment; Compound A group (0.01% Compound A, 0.1% Compound A, 0.25% Compound A, 0.5% Compound A, 2% Compound A and 5% Compound A); Compound B group (0.01% Compound B, 0.1% Compound B, 0.25% Compound B, 0.5% Compound B, 2% Compound B and 5% Compound B); Compound C (0.01% Compound C, 0.1% Compound C, 0.25% Compound C, 0.5% Compound C, 2% Compound C and 5% Compound C); depilatory cream.

[0100] 2. Experimental Methods 2.1 Establishment of Psoriasis Model 2.1.1 Modeling Method and Drug Administration After the adaptive feeding ended, the back skin of the mice was depilated. After anesthetizing the mice, they were placed on an animal fixator to completely expose the back hair area. The back hair of the mice was initially shaved with an electric shaver, and the shaved area was approximately 3 cm × 2 cm. Then, the depilatory cream was thinly applied to the shaved area. After 30 seconds, the depilatory cream and hair were wiped off. After shaving, the status of the mice was observed, and modeling was performed 2 days later under normal feeding. At the same time every morning, 55 mg of 5% imiquimod cream was applied to the exposed parts of the back skin of all the mice in the modeling and drug administration groups. At the same time every afternoon, the corresponding drugs were applied to all the mice: the external application dose of the calcipotriol ointment group was 50 mg, and the application dose of the calcipotriol ointment group was 10 mg; the modeling and drug administration were continued for 5 days.

[0101] 2.1.2 Specimen collection On the 5th day after modeling, that is, the 6th day, the mice in each group were sacrificed by cervical dislocation. The skin tissue of the back skin lesions of the mice was cut off with sterile scissors, the tissue was quickly collected and marked. One part of the tissue was stored in 4% tissue fixative, and the other part was put into a cryotube and frozen in liquid nitrogen for 1 hour and then transferred to a -80 °C water tank for frozen storage for subsequent experiments.

[0102] 2.1.3 Morphological observation On the 1st, 4th, and 6th days of modeling, the back skin conditions of the mice were photographed and recorded to observe the dynamic changes in the skin lesions on the back of the mice, including the rash erythema, scales, and infiltration conditions that appeared in the modeling area: observe the vital signs of the mice in each group, such as reactivity, activity level, mental state, hair color, whether there is scratching of the skin lesions, mortality, etc.

[0103] 2.1.4 PASI score According to the requirements of the PASI score, the changes in the back skin lesions of the mice in each group were statistically analyzed, and the scores were recorded from 0 to 4 points respectively in three aspects: scales, infiltration, and erythema to evaluate the therapeutic effects of the drugs in each group.

[0104] PASI scoring criteria: 0 points = no erythema or residual pigmentation, no scales or flush with the skin; 1 point = mild, showing pink or light erythema, fine scales attached to some skin lesions or the skin lesions slightly protruding above the skin surface; 2 points = moderate, showing darker pink patches, the skin lesion surface is not completely covered with flaky scales or the edge of the skin lesion is moderately raised and easy to touch; 3 points = severe, showing red to dark red patches, almost all the skin lesion surfaces are covered with thick laminated scales or the skin lesions are significantly raised with clear edges; 4 points = extremely severe, showing extremely dark red patches, all the skin lesion surfaces are covered with very thick laminated scales or the skin lesions are extremely thickened and raised with very clear edges; Add the scores of the three items to calculate the total score (total score = scale + infiltration + erythema, 0 - 12 points). The higher the score, the more severe the skin lesion condition, and dynamically evaluate the changes in the skin lesions of mice.

[0105] 2.2 Index detection 2.2.1 HE staining After the treatment ended, the mice in each group were sacrificed by cervical dislocation. The skin at the target skin lesion on the back of the mice was cut with a sterile surgical scissors and cut into pieces of 0.2×0.4 cm in size and placed in a 2 ml centrifuge tube containing 1 ml of 4% tissue cell fixative for fixation. Embed the wax block and make subsequent pathological sections, and perform hematoxylin - eosin (HE) staining to observe the epidermal thickness and tissue inflammation.

[0106] 2.2.2 Fluorescent quantitative PCR detection Take an appropriate amount of skin and place it in a 2 ml RNase - free EP tube. Add 500 μl of Trizol reagent and an appropriate amount of grinding beads to each tube, and quickly homogenize in a tissue homogenizer. After standing until the foam disappears, centrifuge at 12000 r at 4℃ for 15 min, transfer the supernatant to a new RNase - free EP tube, then add 100 μl of chloroform, quickly shake evenly and then stand for 10 min, and centrifuge at 12000 r at 4℃ for 15 min. Slowly aspirate 200 μl of the upper aqueous phase in the EP tube to a new RNase - free EP tube, add 200 μl of isopropanol, invert and mix well, stand for 10 min and then centrifuge at 12000 r at 4℃ for 10 min. Pour out the liquid, invert and dry, then add 200 μl of 75% absolute ethanol to the tube, shake evenly up and down, centrifuge at 12000 r at 4℃ for 5 min, discard the supernatant, and let the precipitate dry at room temperature in a laminar flow hood. Add 20 μl of DEPC water to dissolve the precipitate.

[0107] Use a reverse transcription kit to reverse - transcribe RNA into cDNA. Using GAPDH as an internal reference and SYBR Green as a dye, perform fluorescent quantitative PCR amplification with cDNA as a template. The reaction system is 20 μL, and the amplification conditions are: pre - denaturation at 95℃ for 10 minutes; denaturation at 95℃ for 15 seconds, annealing at 60℃ for 1 minute, 40 amplification cycles; melting curve amplification, 5 seconds at 95℃, 5 seconds at 60℃; extension at 95℃ for 5 seconds. Calculate and compare the differences in the expression of S100A8, S100A9, K6, K16, IL - 17α, IL - 22, IL - 23α, IL - 1α, IL - 1β, CXCL1, CXCL2, VEGF - A, and ICAM - 1 in each group.

[0108] 2.3 Statistical processing Same as Example 1 2.4 Result analysis 2.4.1 Application effects of compound A at different concentrations on the psoriasis mouse model 2.4.1.1 Effects of matrix, compound A (0.01%, 0.1%, 0.25%, 0.5%, 2% and 5%), and calcipotriol on PASI scores of mouse skin in psoriasis cases Psoriasis is a chronic skin disease characterized by red patches on the skin covered with silvery scales. The PASI score is a commonly used method to evaluate the severity of psoriasis. As Figure 1 shown, compared with the IMQ + matrix group, the PASI scores of mice in each drug administration group were decreased to varying degrees (P < 0.05, P < 0.01, P < 0.001). Compared with the IMQ + calcipotriol group, the PASI score of mice with 0.5% compound A was significantly decreased (P < 0.01); among them, the two concentrations of 0.01% and 5% were used to screen the effective concentration of the compound. The results showed that the effect of 0.01% was worse than that of 0.1%, and the effect of 5% was worse than that of 2%. Therefore, for the consideration of optimizing the experimental design and cost - effectiveness, the efficacy of the compound in the concentration range of 0.1% - 2% was mainly explored in the psoriasis mouse model in the follow - up.

[0109] 2.4.1.2 Conditions of changes in the back skin of mice caused by matrix, compound A (0.1%, 0.25%, 0.5% and 2%), and calcipotriol in psoriasis cases Skin condition is an important indicator for judging the severity of psoriasis. The skin of psoriasis patients is mainly manifested as redness, scaling and itching. The therapeutic effect of drugs can be judged according to the severity of the skin surface.

[0110] After modeling, the changes in the skin lesions of mice were observed daily, and different degrees of damage were found in each modeling group. As Figure 2 shown, the skin lesions in the IMQ + matrix group were significantly thickened, and typical scabbing, erythema, infiltration and scales appeared, with obvious elevation and clear boundaries; compared with the IMQ + matrix group, different concentrations of compound A groups had varying degrees of improvement. Among them, 0.5% compound A could significantly reduce the degree of skin redness, the area of scales and the thickening of the skin, and the overall skin was relatively smooth; compared with the compound A group, obvious redness was still visible in the IMQ + calcipotriol group.

[0111] 2.4.1.3 Histopathological experimental comparison of matrix, compound A (0.1%, 0.25%, 0.5% and 2%), and calcipotriol in psoriasis Please refer to Figure 3, The HE staining results showed that: in the IMQ + matrix group, the stratum corneum was significantly thickened, with parakeratosis accompanied by hyperkeratosis, epidermal thickening especially in the stratum spinosum, downward extension of epidermal pegs, and inflammatory cell infiltration in the superficial dermis and papillary layer. The overall manifestation was relatively consistent with the pathological manifestation of psoriasis. The thickening of the stratum corneum in different concentrations of Compound A was significantly alleviated, the epidermis was flat, and the inflammatory cell infiltration decreased, among which 0.5% Compound A showed a significant alleviation; in the calcipotriol group, the thickening of the epidermal stratum corneum was also alleviated to some extent, the downward extension of epidermal pegs decreased, and the number of inflammatory cell infiltration decreased, and the pathological changes were improved.

[0112] 2.4.1.4 mRNA expressions of S100A8, S100A9, K6, and K16 in psoriasis by matrix, Compound A (0.1%, 0.25%, 0.5%, and 2%), and calcipotriol AMPs (S100A8 and S100A9) are secreted by keratinocytes in the psoriatic injury response and are overexpressed in psoriatic skin, promoting the activation of immune cells, angiogenesis, and keratinocyte hyperplasia. Keratin is the main structural intermediate filament protein in keratinocytes and is expressed in a highly specific pattern at different differentiation stages of keratinocytes. The expressions of K6 and K16 represent the highly activated and proliferated state of keratinocytes under pathological conditions.

[0113] It was Figure 4 found that: compared with the IMQ + matrix group, the expression levels of S100A8, S100A9, K6, and K16 in each drug administration group were significantly decreased (P < 0.01, P < 0.001, P < 0.0001), and there were slight differences in the inhibitory effects of different concentrations of Compound A; compared with the IMQ + calcipotriol group, 0.5% Compound A was significantly inhibited in S100A9, K6, and K16 (P < 0.01, P < 0.001, P < 0.0001), indicating that both Compound A and calcipotriol significantly inhibited the abnormal expression and proliferation of inflammatory AMPs and keratinocytes in psoriatic-like skin lesions, and 0.5% Compound A had a better effect. 2.4.1.5 mRNA expressions of IL-17A, IL-22, and IL-23A in psoriasis by matrix, Compound A (0.1%, 0.25%, 0.5%, and 2%), and calcipotriol Skin inflammation and immune regulation play a key role in psoriasis. IL-23 is a key factor in the disease pathogenesis. Psoriatic inflammatory cells activate Th17 cells and T cells that produce IL-17 by releasing IL-23, thereby inducing the expression of a large amount of pro-inflammatory factor IL-17, forming the IL-23 / Th17 / IL-17 immune response pathway.

[0114] As Figure 5As shown, after modeling, there was abnormally high expression in the IMQ + matrix group. Compared with the IMQ + matrix group, the expression levels of IL-17A, IL-22, and IL-23A in each drug administration group were significantly decreased (P < 0.01, P < 0.001, P < 0.0001). At the same time, compared with the IMQ + calcipotriol group, 0.5% compound A could also significantly inhibit the contents of IL-17A, IL-22, and IL-23A (P < 0.001, P < 0.01, P < 0.05), indicating that compound A could significantly inhibit the inflammatory response at the psoriatic-like skin lesion site. 2.4.1.6 mRNA Expression of IL-1α, IL-1β, CXCL1, and CXCL2 in Psoriasis by Matrix, Compound A (0.1%, 0.25%, 0.5%, and 2%), and Calcipotriol Skin inflammation and immune regulation play a key role in psoriasis. In the IL-23 / Th17 / IL-17 immune response pathway, IL-17A targets KCs, promotes activated KCs to release inflammatory and chemotactic factors, participates in the generation of the inflammatory network and amplifies the inflammatory response, among which CXCL1 and CXCL2 tend to activate neutrophils.

[0115] According to Figure 6 As shown, after modeling, there was abnormally high expression in the IMQ + matrix group. Compared with the IMQ + matrix group, the expression levels of compound A at different concentrations were decreased to varying degrees, and 0.5% compound A was significantly better than the IMQ + calcipotriol group (P < 0.05, P < 0.01, P < 0.001), indicating that compound A significantly inhibited the inflammatory response at the psoriatic-like skin lesion site. 2.4.1.7 mRNA Expression of VEGF-A and ICAM-1 in Psoriasis by Matrix, Compound A (0.1%, 0.25%, 0.5%, and 2%), and Calcipotriol VEGF-A is a key factor linking inflammation and angiogenesis. As the most potent angiogenic factor, it has a regulatory effect on endothelial cells and inflammatory cells in patients, thereby inducing angiogenesis and a series of inflammatory responses at psoriatic skin lesions. ICAM-1 is a cell adhesion factor that mediates a series of inflammatory responses.

[0116] As Figure 7As shown, the expression level of VEGF-A in the IMQ + matrix group was significantly increased, promoting angiogenesis at the lesion site. The expression of ICAM-1 was increased, accompanied by an inflammatory response. Compared with the IMQ + matrix group, the relative expression levels of VEGF-A and ICAM-1 of compound A and calcipotriol at different concentrations were significantly decreased (P < 0.01, P < 0.05), inhibiting angiogenesis. Compared with the IMQ + calcipotriol group, 0.5% compound A also had an obvious effect on VEGF-A (P < 0.05, P < 0.01), inhibiting the inflammatory response and playing a role in improving psoriasis-like symptoms.

[0117] 2.4.2 Application effects of compound B at different concentrations on a psoriasis mouse model 2.4.2.1 Effects of matrix, compound B (0.01%, 0.1%, 0.25%, 0.5%, 2% and 5%), and calcipotriol on the PASI score of mice in psoriasis cases As Figure 8 shown, compared with the IMQ + matrix group, the PASI scores of mice in each administration group were decreased to varying degrees (P < 0.05, P < 0.01). The PASI score of 0.5% compound B in mice was equivalent to that of the calcipotriol group; among them, the two concentrations of 0.01% and 5% were used to screen the effective concentration of the compound. The results showed that the effect of 0.01% was worse than that of 0.1%, and the effect of 5% was worse than that of 2%. Therefore, for the consideration of optimizing the experimental design and cost-benefit, the efficacy of the compound in the concentration range of 0.1% - 2% was mainly explored in the psoriasis mouse model subsequently.

[0118] 2.4.2.2 Conditions of changes in the back skin of mice by matrix, compound B (0.1%, 0.25%, 0.5% and 2%), and calcipotriol in psoriasis cases It can be Figure 9 seen that the skin lesions in the IMQ + matrix group were significantly thickened, with typical erythema, infiltration and scales, obvious elevation and clear boundaries; compared with the IMQ + matrix group, different concentrations of compound B had varying degrees of improvement, reducing the erythema, scales and skin thickening of the skin lesions; among them, the effect of the 0.5% compound B group was equivalent to that of the IMQ + calcipotriol group.

[0119] 2.4.2.3 Histopathological experimental comparison of matrix, compound B (0.1%, 0.25%, 0.5% and 2%), and calcipotriol in psoriasis As Figure 10As shown, the HE staining results showed that in the IMQ + matrix group, the stratum corneum was significantly thickened, with parakeratosis accompanied by hyperkeratosis, epidermal thickening, especially in the stratum spinosum, elongation of epidermal pegs, and inflammatory cell infiltration in the upper dermis and papillary layer. The overall manifestation was relatively consistent with the pathological manifestations of psoriasis. In the groups with different concentrations of Compound B, the thickening of the stratum corneum was significantly alleviated, the epidermis was flat, and the inflammatory cell infiltration was reduced. Among them, the effect of 0.5% Compound B was significantly better than that of other concentrations. In the calcipotriol group, the thickening of the epidermal stratum corneum was also alleviated, the downward extension of epidermal pegs was reduced, and the number of inflammatory cell infiltrations was reduced, and the pathological changes were improved.

[0120] 2.4.2.4 mRNA Expressions of S100A8, S100A9, K6, and K16 in Psoriasis by Matrix, Compound B (0.1%, 0.25%, 0.5%, and 2%), and Calcipotriol As Figure 11 shown, compared with the IMQ + matrix group, the expression levels of S100A8, S100A9, K6, and K16 in each administration group were significantly decreased (P < 0.0001, P < 0.001). There was no significant difference in the inhibitory effects of different concentrations of Compound B. Compared with the IMQ + calcipotriol group, there was no significant difference in the inhibitory effect between 0.5% Compound B and calcipotriol, indicating that both 0.5% Compound B and calcipotriol significantly inhibited the abnormal expression and proliferation of inflammatory AMPs and keratinocytes in psoriatic lesions.

[0121] 2.4.2.5 mRNA Expressions of IL-17A, IL-22, and IL-23A in Psoriasis by Matrix, Compound B (0.1%, 0.25%, 0.5%, and 2%), and Calcipotriol As Figure 12 shown, after modeling, the IMQ + matrix group showed abnormally high expression. Compared with the IMQ + matrix group, the expression levels in each administration group were decreased to varying degrees. Among them, the inhibitory effects of 0.5% Compound B and 2% Compound B were the most obvious (P < 0.0001), and the inhibitory degree of 0.5% Compound B group in IL-17A was equivalent to that of the IMQ + calcipotriol group.

[0122] 2.4.2.6 mRNA Expressions of IL-1α, IL-1β, CXCL1, and CXCL2 in Psoriasis by Matrix, Compound B (0.1%, 0.25%, 0.5%, and 2%), and Calcipotriol According to Figure 13As shown, after modeling, there was abnormally high expression in the IMQ + matrix group. Compared with the IMQ + matrix group, the expression levels in the compound B groups at different concentrations were all decreased to varying degrees. Among them, the inhibitory effects of 0.5% compound B and 2% compound B were the most obvious (P < 0.01, P < 0.001, P < 0.0001). Moreover, compared with the IMQ + calcipotriol group, there was no significant difference in the inhibitory effects of the 0.5% compound B group on IL-1α and IL-1β.

[0123] 2.4.2.7 Matrix, compound B (0.1%, 0.25%, 0.5% and 2%), calcipotriol on the mRNA of VEGF-A and ICAM-1 in psoriasis As Figure 14 shown, the expression level of VEGF-A in the IMQ + matrix group was significantly increased, promoting angiogenesis in the skin lesions, and the expression of ICAM-1 was increased, accompanied by an inflammatory response. Compared with the IMQ + matrix group, the relative expression levels of VEGF-A in compound B and calcipotriol at different concentrations were significantly decreased (P < 0.05, P < 0.01). The relative expression levels of ICAM-1 in 0.5% compound B and calcipotriol were significantly decreased (P < 0.05, P < 0.01), inhibiting angiogenesis. Compared with the IMQ + calcipotriol group, there was no obvious difference in VEGF-A for 0.5% compound B, and it also inhibited the inflammatory response, playing a role in improving psoriasis-like symptoms.

[0124] 2.4.3 Application effects of different concentrations of compound C on the psoriasis mouse model 2.4.3.1 Effects of matrix, compound C (0.01%, 0.1%, 0.25%, 0.5%, 2% and 5%), calcipotriol on the PASI score of mice in psoriasis cases As Figure 15 shown, compared with the IMQ + matrix group, the PASI scores of mice in each administration group were all decreased to varying degrees (P < 0.05, P < 0.01). Among them, the two concentrations of 0.01% and 5% were used to screen the effective concentrations of the compound. The results showed that the effect of 0.01% was worse than that of 0.1%, and the effect of 5% was worse than that of 2%. Therefore, for the consideration of optimizing the experimental design and cost-effectiveness, the efficacy of the compound in the concentration range of 0.1% - 2% was mainly explored in the psoriasis mouse model subsequently.

[0125] 2.4.3.2 Conditions of the back skin changes of mice in psoriasis cases with matrix, compound C (0.1%, 0.25%, 0.5% and 2%), calcipotriol As Figure 16As shown, the skin lesions in the IMQ + matrix group were significantly thickened, with typical erythema, infiltration, and scales, prominent elevation, and clear boundaries. Compared with the IMQ + matrix group, different concentrations of Compound C also showed varying degrees of improvement, with alleviated skin lesion symptoms, reduced scales, and less obvious infiltration and thickening. Among them, 0.5% Compound C had a better improvement effect than other concentrations.

[0126] 2.4.3.3 Histopathological experimental comparison of matrix, Compound C (0.1%, 0.25%, 0.5%, and 2%), and calcipotriol in psoriasis Please refer to Figure 17 , and the HE staining results showed that: in the IMQ + matrix group, the stratum corneum was significantly thickened, with parakeratosis accompanied by hyperkeratosis, epidermal thickening especially in the stratum spinosum, downward extension of epidermal pegs, and inflammatory cell infiltration in the superficial dermis and papillary layer, and the overall manifestation was relatively consistent with the pathological manifestation of psoriasis. The thickening of the stratum corneum in different concentration Compound C groups also showed a tendency to be alleviated, the epidermis was relatively flat, and the inflammatory cell infiltration decreased. Among them, 0.5% Compound C had a significant alleviation; in the calcipotriol group, the thickening of the epidermal stratum corneum was also alleviated, the downward extension of epidermal pegs decreased, and the number of inflammatory cell infiltrations decreased, and the pathological changes were improved.

[0127] 2.4.3.4 mRNA expression of S100A8, S100A9, K6, and K16 of matrix, Compound C (0.1%, 0.25%, 0.5%, and 2%), and calcipotriol in psoriasis As Figure 18 shown, compared with the IMQ + matrix group, the expression levels of S100A8, S100A9, K6, and K16 in each administration group were significantly decreased (P < 0.001, P < 0.0001). The inhibitory effect of 0.5% Compound C was the best. Compared with the IMQ + calcipotriol group, the inhibitory effect of 0.5% Compound C on S100A9, K6, and K16 was similar, indicating that both Compound C and calcipotriol significantly inhibited the abnormal expression and proliferation of inflammatory AMPs and keratinocytes in psoriatic lesion sites.

[0128] 2.4.3.5 mRNA expression of IL-17A, IL-22, and IL-23A of matrix, Compound C (0.1%, 0.25%, 0.5%, and 2%), and calcipotriol in psoriasis As Figure 19 shown, after modeling, the IMQ + matrix group had abnormal high expression. Compared with the IMQ + matrix group, the expression levels in each administration group were decreased to varying degrees. Among them, 0.5% Compound C had the most obvious inhibitory effect on IL-17A and IL-22A (P < 0.0001). Compared with the IMQ + calcipotriol group, the effect of Compound C was slightly different.

[0129] 2.4.3.6 Matrix, Compound C (0.1%, 0.25%, 0.5% and 2%), Calcipotriol mRNA expression of IL-1α, IL-1β, CXCL1 and CXCL2 in psoriasis According to Figure 20 As shown, after modeling, the IMQ + matrix group had abnormally high expression. Compared with the IMQ + matrix group, the expression levels of Compound C at different concentrations were all reduced to varying degrees. Among them, 0.5% Compound C had the most obvious inhibitory effect on IL-1α (P < 0.0001), indicating that Compound C significantly inhibited the inflammatory response at the psoriatic lesion site.

[0130] 2.4.3.7 Matrix, Compound C (0.1%, 0.25%, 0.5% and 2%), Calcipotriol mRNA of VEGF-A and ICAM-1 in psoriasis As Figure 21 shown, the expression level of VEGF-A in the IMQ + matrix group was significantly increased, promoting angiogenesis at the lesion site. The expression of ICAM-1 increased, accompanied by an inflammatory response. Compared with the IMQ + matrix group, the relative expression levels of Compound C and Calcipotriol at different concentrations in VEGF-A were all significantly reduced (P < 0.01, P < 0.05), inhibiting angiogenesis and the inflammatory response, and playing a role in improving psoriatic symptoms. Among them, the drug effect of 0.5% Compound C was better than that of other concentrations.

[0131] Example 2 Application of Hesperidin Derivatives in Improving Atopic Dermatitis Mouse Model 1. Experimental Materials 1.1 Experimental Animals Healthy female, SPF clean-grade C57 mice, 6 - 8 weeks old.

[0132] 1.2 Experimental Reagents DNFB; Matrix; Compound A group (0.01% Compound A, 0.1% Compound A, 0.25% Compound A, 0.5% Compound A, 1% Compound A, 3% Compound A and 5% Compound A); Compound B group (0.01% Compound B, 0.1% Compound B, 0.25% Compound B, 0.5% Compound B, 1% Compound B, 3% Compound B and 5% Compound B); Compound C (0.01% Compound C, 0.1% Compound C, 0.25% Compound C, 0.5% Compound C, 1% Compound C, 3% Compound C and 5% Compound C); Eloson Cream; Depilatory Cream; 2. Experimental Methods 2.1 Establishment of Atopic Dermatitis Model 2.1.1 Drug Preparation ① 0.4% DNFB solution: Weigh 1 mL of acetone-olive oil matrix (3:1) and 4 μL of DNFB solution, mix well, store in a cool and dark place for later use.

[0133] ② 0.05% DNFB solution: Weigh 1 mL of acetone-olive oil matrix (3:1) and 0.5 μL of DNFB solution, mix well, store in a cool and dark place for later use.

[0134] 2.1.2 Establishment and administration of atopic dermatitis (AD) model Prepare the model according to the following steps: ① Skin preparation: Before the experiment, remove the abdominal hair of each mouse with an electric shaver, with an area of about 2 cm × 2 cm. ② First sensitization: On day 0 of the experiment, apply 50 μL of 0.4% DNFB to the abdominal skin of all mice for sensitization once. ③ Second challenge and administration: On day 6 of the experiment, remove the dorsal hair of each mouse with an electric shaver, with an area of about 3 cm × 3 cm; on day 7 of the experiment, apply 200 μL of 0.05% DNFB to the dorsal skin of all mice for challenge. 30 minutes later, apply 30 mg of the corresponding group drug to the back, and continue the challenge and administration until day 11.

[0135] 2.1.3 Specimen collection On day 11 of the experiment, euthanize the mice in each group by cervical dislocation. Use sterile scissors to cut the skin tissue at the target skin lesion on the back of the mice, quickly collect the tissue and make good marks, put it into a cryotube and freeze it in liquid nitrogen for 1 hour, and then transfer it to a -80 °C freezer for storage for subsequent experiments.

[0136] 2.1.4 Observation indicators 2.1.4.1 Dermatitis score Refer to the SCORAD scoring standard to observe the dermatitis situation of the dorsal skin of the mice in each group, including erythema, papules or edema, exudation or crusting, epidermal exfoliation, dryness and lichenification. Each clinical symptom is scored according to the grades of 0 points (none), 1 point (mild), 2 points (moderate) and 3 points (severe). Evaluate on day 11 of the experiment and calculate the dermatitis score of each group. The skin lesion inflammation degree scoring table is shown in Table 1.

[0137] Table 1 Skin lesion inflammation degree scoring table

[0138] 2.1.4.2 Observation of scratching behavior of mice Referring to the method of KURAISHI et al., the scratching behavior of mice was observed. On the 11th day of the experiment, the number of times the mice in 14 groups scratched the shaved area was counted. The counting duration for the scratching times of each group of mice was 10 minutes. A continuous scratching of the back by the hind limbs was regarded as 1 scratching activity. When the hind paw landed or was licked, it represented the end of this scratching, and continuous long-term scratching was counted as 1 time.

[0139] 2.2 Index detection 2.2.1 Fluorescent quantitative PCR detection The specific method is the same as that in "Example 1" 2.3 Statistical processing The same as Example 1 2.4 Result analysis 2.4.1 Application effects of compound A at different concentrations on the atopic dermatitis mouse model 2.4.1.1 Effects of the matrix, compound A (0.01%, 0.1%, 0.25%, 0.5%, 1%, 3% and 5%), and Eloson on the dermatitis degree score of the back skin of mice and the number of scratching times in atopic dermatitis cases.

[0140] Clinically, the main symptoms of atopic dermatitis include erythema, pruritus, eczema lesions, edema and skin thickening. In severe cases, there will be fluid exudation and bacterial infection. The dermatitis score and the number of scratching times are important indicators for evaluating the severity of the disease.

[0141] As Figure 22 shown, compared with the DNFB + matrix group, the dermatitis symptom scores and the number of scratching times of the mice in each administration group were significantly reduced (P < 0.05, P < 0.01, P < 0.001, P < 0.0001); compared with the DNFB + Eloson group, the dermatitis score of the mice with 0.5% compound A was significantly reduced (P < 0.05), and the number of scratching times showed a decreasing trend.

[0142] Among them, two concentrations of 0.01% and 5% were used to screen the effective concentration of the compound. The results showed that the effect of 0.01% was relatively poor compared with 0.1%, and the effect of 5% was relatively poor compared with 3%. Therefore, considering the optimization of the experimental design and cost-effectiveness, the efficacy of the compound in the concentration range of 0.1% - 3% was mainly explored in the atopic dermatitis mouse model subsequently.

[0143] 2.4.1.2 mRNA expression of S100A8, S100A9 and K6 in the matrix, compound A (0.1%, 0.25%, 0.5%, 1% and 3%), and Eloson in atopic dermatitis The S100A8 and S100A9 proteins belong to the DAMP molecules. When tissues are damaged and infected, cells in a stressed and dying state can release DAMPs intracellularly, which can act as ligands for some cell surface receptors and induce signals for the production of pro-inflammatory factors, leading to an inflammatory response. In the lesioned areas of AD patients, keratinocytes show abnormal differentiation and abnormal expression of keratin. The expression of K6 related to hyperplasia specificity increases, and the abnormal expression of keratin directly affects the integrity of the epidermal tissue structure, resulting in abnormal skin barrier function.

[0144] As Figure 23 shown, compared with the DNFB + matrix group, the expression levels of S100A8 and S100A9 in each drug administration group were significantly decreased (P<0.01, P<0.001, P<0.0001). There were slight differences in the inhibitory effects of compound A at different concentrations. The expression levels of 0.5% compound A and Eloson in K6 also decreased (P<0.05, P<0.01); compared with the DNFB + Eloson group, 0.5% compound A significantly inhibited both in S100A8 and S100A9 (P<0.01, P<0.001), and although the expression level of K6 increased, there was no significant difference, thus promoting the recovery of the skin epidermal barrier.

[0145] 2.4.1.3 mRNA expression of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in atopic dermatitis by matrix, compound A (0.1%, 0.25%, 0.5%, 1%, and 3%), and Eloson IL-1α and IL-1β are the main pro-inflammatory cytokines in the body. IL-4 and IL-13 are considered to be the marker cytokines of Th2 cells and early atopic dermatitis. IL-4 and IL-13 can also regulate the epidermal barrier and inhibit the production of antimicrobial peptides.

[0146] As Figure 24 shown, compared with the DNFB + matrix group, the expression levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in each drug administration group were significantly decreased (P<0.05, P<0.01, P<0.001, P<0.0001), improving the skin inflammatory response; and compared with the Eloson group, 0.5% compound A had a significant inhibitory effect on the expression of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 (P<0.05, P<0.01).

[0147] 2.4.1.4 mRNA expression of IL-17A, IL-22, and IL-23A in atopic dermatitis by matrix, compound A (0.1%, 0.25%, 0.5%, 1%, and 3%), and Eloson IL-17 is a newly discovered pro-inflammatory cytokine mainly secreted by helper T cells 17. A large number of Th17 cells infiltrate the skin lesions of AD patients, and stimulate the abnormal activation of keratinocytes by secreting cytokines such as IL17 and IL-23, showing an inflammatory amplification effect.

[0148] As Figure 25 shown, compared with the DNFB + matrix group, the expression levels of IL-17A, IL-22, and IL-23A in each drug administration group were significantly decreased (P < 0.05, P < 0.01, P < 0.001, P < 0.0001), improving the skin inflammatory response; and compared with the Eloson group, the expression of 0.5% compound A in IL-17A, IL-22, and IL-23A was significantly inhibited (P < 0.05, P < 0.01).

[0149] 2.4.1.5 mRNA expression of VEGF-A and ICAM-1 in atopic dermatitis by matrix, compound A (0.1%, 0.25%, 0.5%, 1%, and 3%), and Eloson AD is a chronic and recurrent inflammatory skin disease. Chronic inflammation will inevitably involve the problem of angiogenesis to facilitate the infiltration of inflammatory cells and the chronic course.

[0150] As Figure 26 shown, compared with the DNFB + matrix group, the expression levels of VEGF-A and ICAM-1 in each drug administration group were significantly decreased (P < 0.01, P < 0.001), inhibiting angiogenesis and reducing vascular permeability; compared with the Eloson group, the expression of 0.5% compound A in VEGF-A and ICAM-1 was significantly inhibited (P < 0.05).

[0151] 2.4.2 Application effects of different concentrations of compound B on a mouse model of atopic dermatitis 2.4.2.1 Effects of matrix, compound B (0.01%, 0.1%, 0.25%, 0.5%, 1%, 3%, and 5%), and Eloson on the dermatitis degree score of the back skin of mice and the scratching times of mice in atopic dermatitis cases As Figure 27 shown, compared with the DNFB + matrix group, the mice in the 0.25% and 0.5% compound B and Eloson groups had significantly lower dermatitis scores (P < 0.05, P < 0.01, P < 0.001), and the scratching times of mice in each drug administration group were significantly reduced (P < 0.001, P < 0.0001); compared with the DNFB + Eloson group, there was no significant difference in the scratching times of the mice in the 0.5% compound B group.

[0152] Among them, two concentrations of 0.01% and 5% were used to screen the effective concentration of the compound. The results showed that the effect of 0.01% was worse than that of 0.1%, and the effect of 5% was worse than that of 3%. Therefore, considering the optimization of experimental design and cost-effectiveness, the efficacy of the compound in the concentration range of 0.1% - 3% was mainly explored in the atopic dermatitis mouse model subsequently.

[0153] 2.4.2.2 mRNA expression of S100A8, S100A9 and K6 in atopic dermatitis with matrix, compound B (0.1%, 0.25%, 0.5%, 1% and 3%), and Eloson As Figure 28 shown, compared with the DNFB + matrix group, the expression levels of S100A8 and S100A9 in each administration group were significantly decreased (P < 0.01, P < 0.001, P < 0.0001). There were slight differences in the inhibitory effects of compound B at different concentrations. The expression levels of 0.5% and 1% compound B and Eloson in K6 were also significantly decreased (P < 0.05, P < 0.01). Although the expression levels of compound B at other concentrations increased, there were no obvious differences. Compared with the DNFB + Eloson group, the reduction effects of the expression levels of S100A8 and S100A9 of 0.5% compound B were comparable, thus promoting the recovery of the skin epidermal barrier.

[0154] 2.4.2.3 mRNA expression of IL-1α, IL-1β, IL-4, IL-13, CXCL1 and CXCL2 in atopic dermatitis with matrix, compound B (0.1%, 0.25%, 0.5%, 1% and 3%), and Eloson As Figure 29 shown, compared with the DNFB + matrix group, the expression levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1 and CXCL2 in each administration group were significantly decreased (P < 0.05, P < 0.01, P < 0.001, P < 0.0001); compared with the Eloson group, the expression effects of 0.5% compound B in IL-1α, IL-1β, IL-4 and IL-13 were comparable.

[0155] 2.4.2.4 mRNA expression of IL-17A, IL-22 and IL-23A in atopic dermatitis with matrix, compound B (0.1%, 0.25%, 0.5%, 1% and 3%), and Eloson As Figure 30As shown, compared with the DNFB + matrix group, the expression levels of IL-22 and IL-23A in each administration group were significantly decreased (P < 0.01, P < 0.001, P < 0.0001). In IL-17A, 0.25%, 0.5%, 1% Compound B and Eloson could significantly decrease it (P < 0.05, P < 0.01, P < 0.001); compared with the Eloson group, the expression effects of 0.5% Compound B in IL-22 and IL-23A were equivalent. 2.4.2.5 mRNA expression of VEGF-A and ICAM-1 in atopic dermatitis by matrix, Compound B (0.1%, 0.25%, 0.5%, 1% and 3%), and Eloson As Figure 31 shown, compared with the DNFB + matrix group, the expression levels of VEGF-A and ICAM-1 in each administration group were significantly decreased (P < 0.05, P < 0.01, P < 0.001), inhibiting angiogenesis and reducing vascular permeability; compared with the Eloson group, the expression effects of 0.5% and 1% Compound B in VEGF-A and ICAM-1 were equivalent.

[0156] 2.4.3 Application effects of different concentrations of Compound C on the atopic dermatitis mouse model 2.4.3.1 Effects of matrix, Compound C (0.01%, 0.1%, 0.25%, 0.5%, 1%, 3% and 5%), and Eloson on the dermatitis degree score of the back skin of mice and the scratching times of mice in atopic dermatitis cases.

[0157] As Figure 32 shown, compared with the DNFB + matrix group, the mice in the 0.25%, 0.5%, 1% Compound C and Eloson groups had significantly decreased dermatitis scores (P < 0.05, P < 0.01, P < 0.001), and the scratching times of each administration group of mice were significantly reduced (P < 0.001, P < 0.0001); compared with the DNFB + Eloson group, the inhibitory effects of Compound C on the dermatitis score and scratching times of mice were slightly lower than those of Eloson. 2.4.3.2 mRNA expression of S100A8, S100A9 and K6 in atopic dermatitis by matrix, Compound C (0.1%, 0.25%, 0.5%, 1% and 3%), and Eloson Among them, two concentrations of 0.01% and 5% were used to screen the efficacy concentration of the compound. The results showed that the effect of 0.01% was worse than that of 0.1%, and the effect of 5% was worse than that of 3%. Therefore, for the consideration of optimizing the experimental design and cost - benefit, the efficacy of the compound in the concentration range of 0.1% - 3% was mainly explored in the atopic dermatitis mouse model subsequently.

[0158] As Figure 33As shown, compared with the DNFB + matrix group, the expression levels of S100A8 and S100A9 in each administration group were significantly decreased (P<0.01, P<0.001, P<0.0001). There were slight differences in the inhibitory effects of compound C at different concentrations, and the expression of compound C increased in K6. Compared with the DNFB + Eloson group, the reduction effects of the expression levels of S100A8 and S100A9 in 0.5% compound C were comparable, thereby promoting the recovery of the skin epidermal barrier.

[0159] 2.4.3.3 mRNA expression of IL-1α, IL-1β, IL-4, IL-13, CXCL1 and CXCL2 in atopic dermatitis by matrix, compound C (0.1%, 0.25%, 0.5%, 1% and 3%), and Eloson As Figure 34 shown, compared with the DNFB + matrix group, the expression levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1 and CXCL2 in each administration group were significantly decreased or showed an obvious decreasing trend (P<0.05, P<0.01, P<0.001, P<0.0001); compared with the Eloson group, the expression effects of 0.5% compound C in IL-1β, IL-4 and IL-13 were comparable.

[0160] 2.4.3.4 mRNA expression of IL-17A, IL-22 and IL-23A in atopic dermatitis by matrix, compound C (0.1%, 0.25%, 0.5%, 1% and 3%), and Eloson As Figure 35 shown, compared with the DNFB + matrix group, the expression levels of IL-22 and IL-23A in each administration group were significantly decreased (P<0.05, P<0.01, P<0.001, P<0.0001), and there was a decreasing trend in IL-17A; compared with the Eloson group, the expression effects of 0.5% compound C in IL-22 and IL-23A were comparable.

[0161] 2.4.3.5 mRNA expression of VEGF-A and ICAM-1 in atopic dermatitis by matrix, compound C (0.1%, 0.25%, 0.5%, 1% and 3%), and Eloson.

[0162] As Figure 36 shown, compared with the DNFB + matrix group, the expression levels of VEGF-A and ICAM-1 in each administration group were significantly decreased (P<0.05, P<0.01, P<0.001), inhibiting angiogenesis and reducing vascular permeability; compared with the Eloson group, the expression effect of 0.5% compound C in VEGF-A was comparable.

[0163] Example 3 Application of Hesperidin Derivatives in Improving a Mouse Model of Sensitive Skin (Barrier Damage) 1. Experimental Materials 1.1 Experimental Animals Healthy female, SPF clean-grade C57 mice, 6 - 8 weeks old.

[0164] 1.2 Experimental Reagents Tape; matrix; calcipotriol, compound A groups (0.01% compound A, 0.1% compound A, 0.25% compound A, 0.5% compound A, 1% compound A, 3% compound A, and 5% compound A); compound B groups (0.01% compound B, 0.1% compound B, 0.25% compound B, 0.5% compound B, 1% compound B, 3% compound B, and 5% compound B); compound C groups (0.01% compound C, 0.1% compound C, 0.25% compound C, 0.5% compound C, 1% compound C, 3% compound C, and 5% compound C); depilatory cream 2. Experimental Methods 2.1 Establishment of a Mouse Model of Sensitive Skin 2.1.1 Modeling Method and Drug Administration Remove the hair on the back of the mice. The next day, use Tape to peel the back skin of the mice 3 times. After 30 minutes of Tape action, apply 30 mg of the corresponding drug respectively, once in the morning and once in the evening, and continuously perform Tape peeling and application for three days.

[0165] 2.1.2 Specimen Collection On the 4th day of the experiment, sacrifice the mice in each group by cervical dislocation. Use sterile scissors to cut the skin tissue at the skin lesion on the back of the mice, quickly collect the tissue and make good marks, put it into a cryotube and freeze it in liquid nitrogen for 1 hour, and then transfer it to a -80°C water tank for freezing preservation for subsequent experiments.

[0166] 2.2 Index Detection 2.2.1 Detection of TEWL Value of Mouse Skin On the 1st day before modeling and the 4th day of the experiment, use a VapoMeter to measure the skin on the neck and back of the mice. The measurement is carried out indoors without direct sunlight, at a room temperature of 20 - 25°C and a relative air humidity of 10% - 60%. When measuring, fix the mice well to ensure that the measuring instrument is in perpendicular contact with the surface of the skin to be measured. Measure each test site 3 times and take the average of the 3 values.

[0167] 2.2.2 Fluorescent Quantitative PCR Detection The specific method is the same as that in "Example 1" 2.3 Statistical Processing The same as Example 1 2.4 Result Analysis 2.4.1 Application effects of Compound A at different concentrations on a sensitive skin mouse model 2.4.1.1 Effects of matrix, Compound A (0.01%, 0.1%, 0.25%, 0.5%, 1%, 3% and 5%), and calcipotriol on the TEWL value of the dorsal skin of mice in sensitive skin cases TEWL indicates the water loss situation by measuring the water vapor pressure gradient on the skin surface and is a classic evaluation index for skin barrier function. When the skin barrier function is impaired, the skin's water retention function weakens, water loss increases, and the TEWL value increases. The change in the TEWL value can accurately and stably reflect the early recovery process of the skin barrier.

[0168] As Figure 37 shown, before modeling, there was no significant change in the TEWL value of the skin of mice in all groups; on the 4th day after modeling and administration, the TEWL values of the skin of mice in all groups increased to varying degrees, and the increase in the Tape + matrix group was significant, indicating that the mechanical barrier function of the mice's skin was damaged; compared with the Tape + matrix group, the TEWL values of the skin of mice in each administration group were significantly decreased (P < 0.01, P < 0.001), and compared with the Tape + calcipotriol group, 0.5% Compound A was significantly decreased (P < 0.05), and the skin barrier recovery effect was significant.

[0169] Among them, two concentrations of 0.01% and 5% were used to screen the efficacy concentration of the compound. The results showed that 0.01% was less effective than 0.1%, and 5% was less effective than 3%. Therefore, for the consideration of optimizing the experimental design and cost - effectiveness, the efficacy of the compound in the concentration range of 0.1% - 3% was focused on exploring in the sensitive skin mouse model in the follow - up.

[0170] 2.4.1.2 mRNA expression of S100A8, S100A9, K6 and K16 of matrix, Compound A (0.1%, 0.25%, 0.5%, 1% and 3%), and calcipotriol in sensitive skin cases Keratin K6 and K16 are crucial for the epidermal permeability barrier function and the hydration of the stratum corneum, and the calcium - binding proteins S100A8 and S100A9 can induce the synthesis of other skin proteins and sebum.

[0171] As Figure 38As shown, the mRNA levels of S100A8, S100A9, K6, and K16 were significantly increased after Tape treatment, indicating that Tape treatment disrupted the epidermal barrier function, thus causing the generation of sensitive skin to a certain extent. After application of each drug administration group, the increase of the above factors was also significantly inhibited (P<0.001, P<0.0001), and compared with the Tape+calcipotriol group, 0.5% Compound A decreased significantly (P<0.05), indicating that Compound A can promote the recovery of the epidermal barrier function of sensitive skin and inhibit epidermal hyperplasia of sensitive skin.

[0172] 2.4.1.3 mRNA expression of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in cases of sensitive skin by matrix, Compound A (0.1%, 0.25%, 0.5%, 1%, and 3%), and calcipotriol As Figure 39 shown, after Tape treatment, the mRNA levels of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in skin tissues were significantly increased. Different concentrations of Compound A could inhibit the increase of the mRNA levels of the above inflammatory factors in skin tissues to a certain extent (P<0.01, P<0.001, P<0.0001). Compared with the Tape+calcipotriol group, 0.5% Compound A decreased significantly (P<0.05, P<0.01), which has a regulatory effect on the expression levels of inflammatory factors, and thus can effectively improve skin inflammation of sensitive skin.

[0173] 2.4.1.4 mRNA expression of IL-17A, IL-22, and IL-23A in cases of sensitive skin by matrix, Compound A (0.1%, 0.25%, 0.5%, 1%, and 3%), and calcipotriol IL-23 / Th17 is a key pathway for immune regulation of skin diseases. IL-23 is secreted by skin-resident dendritic cells and induces the polarization and production of Th17 cells (such as IL-17A / IL-22, etc.) of pro-inflammatory mediators.

[0174] As Figure 40 shown, after Tape treatment, the mRNA expression levels of immune cell activity-related factors IL-17A and IL-22A were significantly increased. Different concentrations of Compound A could inhibit the increase of the mRNA levels of the above immune factors in skin tissues to a certain extent (P<0.01, P<0.001). Compared with the Tape+calcipotriol group, 0.25% and 0.5% Compound A decreased significantly in IL-17A, IL-22, and IL-23A (P<0.05, P<0.01), which has a regulatory effect on the expression levels of immune activity-related factors of sensitive skin, and thus improves the immune function of sensitive skin.

[0175] 2.4.1.5 mRNA Expression of VEGF-A and ICAM-1 in Sensitive Skin Cases with Matrix, Compound A (0.1%, 0.25%, 0.5%, 1% and 3%), and Calcipotriol Superficial microvascular hyperplasia and high permeability phenotypes are important features in the occurrence and development of sensitive skin. As Figure 41 shown, after Tape treatment, the mRNA expression levels of the vascular permeability-related factors VEGF-A and ICAM-1 increased significantly. Different concentrations of Compound A could inhibit the increase in the mRNA levels of the above factors in skin tissues to a certain extent (P<0.01, P<0.001). Compared with the Tape + Calcipotriol group, 0.5% Compound A significantly decreased VEGF-A and ICAM-1 (P<0.05), regulated the expression levels of vascular permeability factors, and thus could effectively improve the vascular permeability of sensitive skin.

[0176] 2.4.2 Application Effects of Different Concentrations of Compound B on a Mouse Model of Sensitive Skin 2.4.2.1 Effects of Matrix, Compound B (0.01%, 0.1%, 0.25%, 0.5%, 1%, 3% and 5%), and Calcipotriol on the TEWL Value of Mouse Back Skin in Sensitive Skin Cases As Figure 42 shown, before modeling, there were no significant changes in the TEWL values of the skin of mice in all groups; on the 4th day after modeling and administration, the TEWL values of the skin of mice in all groups increased to varying degrees, and the increase was significant in the Tape + Matrix group, indicating that the mechanical barrier function of the mouse skin was damaged; compared with the Tape + Matrix group, the TEWL values of the skin of mice in each administration group also decreased significantly (P<0.05, P<0.01, P<0.001). Compared with the Tape + Calcipotriol group, the skin barrier recovery effect of 0.5% Compound B was comparable.

[0177] Among them, two concentrations of 0.01% and 5% were used to screen the effective concentrations of the compound. The results showed that the effect of 0.01% was worse than that of 0.1%, and the effect of 5% was worse than that of 3%. Therefore, for the consideration of optimizing the experimental design and cost-effectiveness, the efficacy of the compound in the concentration range of 0.1% - 3% was mainly explored in the mouse model of sensitive skin subsequently.

[0178] 2.4.2.2 mRNA Expression of S100A8, S100A9, K6 and K16 in Sensitive Skin Cases with Matrix, Compound B (0.1%, 0.25%, 0.5%, 1% and 3%), and Calcipotriol As Figure 43As shown, the mRNA levels of S100A8, S100A9, K6, and K16 were significantly increased after Tape treatment, indicating that Tape treatment disrupted the epidermal barrier function, thus causing the generation of sensitive skin to a certain extent. After application in each drug administration group, the increase of the above factors was significantly inhibited (P<0.01, P<0.001, P<0.0001), and compared with the Tape+calcipotriol group, the effects of 0.5% and 1% Compound B were equivalent, indicating that it has a certain effect on the restoration of the epidermal barrier function of sensitive skin.

[0179] 2.4.2.3 mRNA expression of IL-1α, IL-1β, IL-4, IL-13, CXCL1, and CXCL2 in sensitive skin cases by matrix, Compound B (0.1%, 0.25%, 0.5%, 1%, and 3%), and calcipotriol As Figure 44 shown, after Tape treatment, the mRNA levels of IL-1α, IL-1β, IL-4, IL-13, and CXCL1 in skin tissues were significantly increased. Compound B could inhibit the increase of the mRNA levels of the above inflammatory factors in skin tissues to a certain extent (P<0.01, P<0.001, P<0.0001). Compared with the Tape+calcipotriol group, the effects of 0.5% and 1% Compound B were equivalent in IL-1α, CXCL1, and CXCL2, and had the effect of improving skin inflammation in sensitive skin.

[0180] 2.4.2.4 mRNA expression of IL-17A, IL-22, and IL-23A in sensitive skin cases by matrix, Compound B (0.1%, 0.25%, 0.5%, 1%, and 3%), and calcipotriol As Figure 45 shown, after Tape treatment, the mRNA expression levels of the immune cell activity-related factors IL-17A, IL-22A, and IL-23A were significantly increased. Compound B had a significant effect on inhibiting the increase of the mRNA levels of the above immune factors in skin tissues (P<0.01, P<0.001). Compared with the Tape+calcipotriol group, the effects of 0.5% and 1% Compound B were equivalent in reducing the expression levels of IL-17A, IL-22A, and IL-23A, and had a certain function of improving the immunity of sensitive skin.

[0181] 2.4.2.5 mRNA expression of VEGF-A and ICAM-1 in sensitive skin cases by matrix, Compound B (0.1%, 0.25%, 0.5%, 1%, and 3%), and calcipotriol As Figure 46As shown, after Tape treatment, the mRNA expression levels of vascular permeability-related factors VEGF-A and ICAM-1 were significantly increased. Compound B could significantly reduce the expression level of VEGF-A in skin tissue (P<0.01, P<0.001), and significantly reduce or show a trend of reducing the expression level of ICAM-1 (P<0.05, P<0.01). Compared with the Tape+calcipotriol group, the expression effects of 0.5% Compound B on VEGF-A and ICAM-1 were comparable, reducing the vascular permeability of sensitive skin.

[0182] 2.4.3 Application effects of different concentrations of Compound C on a mouse model of sensitive skin 2.4.3.1 Effects of matrix, Compound C (0.01%, 0.1%, 0.25%, 0.5%, 1%, 3% and 5%), and calcipotriol on the TEWL value of the dorsal skin of mice in cases of sensitive skin As Figure 47 shown, before modeling, there were no obvious changes in the TEWL values of the skin of mice in all groups; on the 4th day after modeling and administration, the TEWL values of the skin of mice in all groups increased to varying degrees, among which the increase in the Tape+matrix group was obvious, indicating that the mechanical barrier function of the mouse skin was damaged; compared with the Tape+matrix group, the TEWL values of the skin of mice in each administration group were also significantly reduced (P<0.05, P<0.01, P<0.001), and the effect of 0.5% Compound C was the best.

[0183] Among them, two concentrations of 0.01% and 5% were used to screen the effective concentration of the compound. The results showed that the effect of 0.01% was relatively poor compared with 0.1%, and the effect of 5% was relatively poor compared with 3%. Therefore, for the consideration of optimizing the experimental design and cost-benefit, the compound efficacy in the concentration range of 0.1% - 3% was mainly explored in the mouse model of sensitive skin subsequently.

[0184] 2.4.3.2 mRNA expression of S100A8, S100A9, K6 and K16 in matrix, Compound C (0.1%, 0.25%, 0.5%, 1% and 3%), and calcipotriol in cases of sensitive skin As Figure 48 shown, after Tape treatment, there were obvious increases in the mRNA of S100A8, S100A9, K6, and K16, indicating that Tape treatment damaged the epidermal barrier function, thus causing the generation of sensitive skin to a certain extent. After applying Compound C, it could inhibit the increase of the above factors (P<0.01, P<0.001, P<0.0001). Compared with the Tape+calcipotriol group, the inhibitory effect of 0.5% Compound C was comparable, indicating that it had a certain effect on the recovery of the epidermal barrier function of sensitive skin.

[0185] 2.4.3.3 mRNA Expression of Matrix, Compound C (0.1%, 0.25%, 0.5%, 1% and 3%), Calcipotriol for IL-1α, IL-1β, IL-4, IL-13, CXCL1 and CXCL2 in Sensitive Skin Cases As Figure 49 shown, after Tape treatment, the mRNA of IL-1α, IL-1β, IL-4, IL-13, CXCL1 in skin tissue increased significantly. Compound C could inhibit the increase in the mRNA levels of the above inflammatory factors in skin tissue to a certain extent (P<0.05, P<0.01, P<0.001, P<0.0001). Compared with the Tape + Calcipotriol group, 0.5% Compound C had comparable expression effects on IL-1α, IL-1β, CXCL1, CXCL, and improved the skin inflammation of sensitive skin.

[0186] 2.4.3.4 mRNA Expression of Matrix, Compound C (0.1%, 0.25%, 0.5%, 1% and 3%), Calcipotriol for IL-17A, IL-22 and IL-23A in Sensitive Skin Cases As Figure 50 shown, after Tape treatment, the mRNA expression levels of IL-17A, IL-22A and IL-23A increased significantly. Compound C inhibited the increase in the mRNA levels of the above immune factors in skin tissue (P<0.05, P<0.01, P<0.001). Compared with the Tape + Calcipotriol group, 0.5% Compound C had comparable effects on reducing the expression of IL-23A and could improve the immune function of sensitive skin.

[0187] 2.4.3.5 mRNA Expression of Matrix, Compound C (0.1%, 0.25%, 0.5%, 1% and 3%), Calcipotriol for VEGF-A, ICAM-1 in Sensitive Skin Cases As Figure 51 shown, after Tape treatment, the mRNA expression levels of the vascular permeability-related factors VEGF-A and ICAM-1 increased significantly. 0.5% Compound C significantly reduced the mRNA levels of the above factors in skin tissue or showed a trend (P<0.05, P<0.01), reducing the vascular permeability of sensitive skin.

[0188] Example 4 Application of Hesperidin Derivatives in Improving the Mouse Model of Irritant Dermatitis 1. Experimental Materials 1.1 Experimental Animals Healthy female, SPF clean-grade C57 mice, 6 - 8 weeks old.

[0189] 1.2 Experimental Reagents Phorbol-12-myristate-13-acetate (TPA); matrix; compound A group (0.01% compound A, 0.1% compound A, 0.25% compound A, 0.5% compound A, 1% compound A, 3% compound A, and 5% compound A); compound B group (0.01% compound B, 0.1% compound B, 0.25% compound B, 0.5% compound B, 1% compound B, 3% compound B, and 0.5% compound B); compound C group (0.01% compound C, 0.1% compound C, 0.25% compound C, 0.5% compound C, 1% compound C, 3% compound C, and 0.5% compound C); depilatory cream; positive drug (Eloson cream).

[0190] 2. Experimental methods 2.1 Establishment of a TPA-induced irritant dermatitis model in mice 2.1.1 Modeling method and drug administration A mouse irritant dermatitis model was made by topical application of TPA: Every morning, 10 μl of TPA was applied to both sides of the ears of the mice. 30 minutes later, 20 mg of the matrix was applied to both sides of the left ear, and 20 mg of the corresponding drug was applied to both sides of the right ear. At the same time, the drug was applied again every night for 3 consecutive days.

[0191] 2.1.2 Specimen collection On the 4th day of the experiment, the mice in each group were sacrificed by cervical dislocation. The skin tissue of the damaged area on the back of the mice was cut off with sterile scissors. The tissue was quickly collected and labeled, placed in a cryotube, frozen in liquid nitrogen for 1 hour, and then transferred to an -80°C freezer for storage for subsequent experiments.

[0192] 2.2 Index detection 2.2.1 Detection of ear margin thickness The ear margin thickness was recorded with a vernier caliper on the 1st and 4th days of the experiment.

[0193] 2.2.2 Fluorescent quantitative PCR detection The specific method was the same as that in "Example 1". 2.3 Statistical processing The same as Example 1 2.4 Result analysis 2.4.1 Application effects of different concentrations of compound A on the irritant dermatitis mouse model 2.4.1.1 Effects of the matrix, compound A (0.01%, 0.1%, 0.25%, 0.5%, 1%, 3%, and 5%), and positive drug on the ear margin thickness of mice in irritant dermatitis cases TPA is the main active ingredient of croton oil. The TPA-induced mouse irritant dermatitis model is a classic animal model of chemical contact dermatitis. After applying phorbol ester to the auricles of mice, it can cause the release of inflammatory mediators (such as histamine and kinins, etc.) in the ear tissues of mice, leading to vasodilation of the ear tissues, enhanced capillary permeability, and further recruitment of a large number of inflammatory cells to the ear tissues, resulting in inflammatory cell infiltration and ear inflammatory edema.

[0194] As Figure 52 shown, after local application of TPA to the mouse ears, morphological changes in the mouse ears can be seen with the naked eye. In the TPA + matrix group, the ears showed redness, thickening, and edema. Through ear measurement, before the experiment, there was no difference in the thickness of the ear margins among the groups. After modeling, the thickness of the ear margins of all groups of mice increased to varying degrees, and the increase was significant in the TPA + matrix group, indicating the occurrence of inflammatory edema in the mouse ears; compared with the TPA + matrix group, the thickness of the ear margins in each drug administration group decreased significantly (P < 0.01, P < 0.001). Compared with the TPA + positive drug group, 0.5% compound A had a significant inhibitory effect (P < 0.05), reducing ear edema.

[0195] Among them, two concentrations of 0.01% and 5% were used to screen the effective concentrations of the compounds. The results showed that the effect of 0.01% was relatively poor compared with 0.1%, and the effect of 5% was relatively poor compared with 3%. Therefore, considering the optimization of the experimental design and cost - effectiveness, the efficacy of compounds in the concentration range of 0.1% - 3% was mainly explored in the irritant dermatitis mouse model in the follow - up.

[0196] 2.4.1.2 mRNA expression of S100A8, S100A9, and K6 in the irritant dermatitis mice with matrix, compound A (0.1%, 0.25%, 0.5%, 1%, and 3%), and positive drug S100A8, S100A9, and K6 are factors related to the epidermal barrier function. As Figure 53 shown, the mRNA of S100A8, S100A9, and K6 increased significantly after TPA treatment, indicating that TPA treatment damaged the epidermal barrier function, thus causing ear inflammation to a certain extent. After application by the drug administration groups, the increase of S100A8, S100A9, and K6 factors could be significantly inhibited (P < 0.05, P < 0.01, P < 0.001, P < 0.0001). Compared with the TPA + positive drug group, 0.5% compound A could significantly inhibit the expression of S100A8, S100A9, and K6 factors (P < 0.05, P < 0.01), reducing ear inflammation.

[0197] 2.4.1.3 mRNA Expression of Matrix, Compound A (0.1%, 0.25%, 0.5%, 1% and 3%), and Positive Drug for IL-1α, IL-1β, IL-4, IL-13, CXCL1 and CXCL2 in Mice with Irritant Dermatitis IL-1α, IL-1β, IL-4, and IL-13 are the main mediators of inflammatory and immune skin diseases. Both inflammation and immune stimulation can induce the production and secretion of inflammatory factors in keratinocytes. As Figure 54 shown, after TPA treatment, the mRNA expression levels of IL-1α, IL-1β, IL-4, IL-13, and CXCL2 in skin tissues increased. Compound A could significantly inhibit the increase of the above genes to a certain extent (P<0.05, P<0.01, P<0.001, P<0.0001). Compared with the TPA+positive drug group, 0.5% Compound A could reduce the expression of IL-1α, IL-1β, IL-4, IL-13, and CXCL2 factors (P<0.05, P<0.01), showing a local anti-inflammatory effect on ear inflammation.

[0198] 2.4.1.4 mRNA Expression of Matrix, Compound A (0.1%, 0.25%, 0.5%, 1% and 3%), and Positive Drug for IL-17A, IL-22 and IL-23A in Mice with Irritant Dermatitis The IL-23 / IL-17 is a key pathway for immune regulation of skin diseases. IL-17A can significantly induce the synthesis and accumulation of other inflammatory mediators and inflammatory cytokines in target tissues, leading to inflammatory reactions and tissue damage. As Figure 55 shown, after TPA treatment, the mRNA expression levels of IL-17A, IL-22, and IL-23A in skin tissues increased. Compound A could significantly reduce the expression of IL-17A, IL-22, and IL-23A to a certain extent (P<0.01, P<0.001). Compared with the TPA+positive drug group, 0.5% Compound A could reduce the expression of the above factors (P<0.05, P<0.01), showing an improvement effect on the immune function of the ear.

[0199] 2.4.1.5 mRNA Expression of Matrix, Compound A (0.1%, 0.25%, 0.5%, 1% and 3%), and Positive Drug for VEGF-A and ICAM-1 in Mice with Irritant Dermatitis As Figure 56As shown, after TPA treatment, the mRNA expression level of the vascular permeability-related factor VEGF-A increased. Compound A could significantly reduce the expression of VEGF-A in skin tissues (P<0.05, P<0.01), but could increase the expression of ICAM-1. Compared with the TPA + positive drug group, 0.5% Compound A could significantly reduce the expression of the VEGF-A factor (P<0.01).

[0200] 2.4.2 Application effects of different concentrations of Compound B on the mouse model of irritant dermatitis 2.4.2.1 Effects of the matrix, Compound B (0.01%, 0.1%, 0.25%, 0.5%, 1%, 3% and 5%), and the positive drug on the ear margin thickness of mice in cases of irritant dermatitis As Figure 57 shown, after local application of TPA to the mouse ears, morphological changes in the mouse ears were visible to the naked eye. In the TPA + matrix group, the ears showed redness, thickening, and edema. Through ear measurement, before the experiment, there were no differences in the ear margin thickness among the groups. After modeling, the ear margin thickness of all groups of mice increased to varying degrees, and the increase was significant in the TPA + matrix group, indicating the occurrence of inflammatory edema in the mouse ears; compared with the TPA + matrix group, the ear margin thickness of each drug administration group decreased significantly (P<0.05, P<0.01, P<0.001). Compared with the TPA + positive drug group, the inhibitory effect of 0.5% Compound B was comparable, and the ear edema was alleviated.

[0201] Among them, two concentrations of 0.01% and 5% were used to screen the effective concentrations of the compound. The results showed that the effect of 0.01% was relatively poor compared with 0.1%, and the effect of 5% was relatively poor compared with 3%. Therefore, for the consideration of optimizing the experimental design and cost-effectiveness, the compound efficacy in the concentration range of 0.1% - 3% was mainly explored in the mouse model of irritant dermatitis in the follow-up.

[0202] 2.4.2.2 mRNA expression of S100A8, S100A9, and K6 in the matrix, Compound B (0.1%, 0.25%, 0.5%, 1%, and 3%), and the positive drug in mice with irritant dermatitis As Figure 58 shown, the mRNA of S100A8 and S100A9 after TPA treatment increased significantly, indicating that TPA treatment damaged the epidermal barrier function, thus causing ear inflammation to a certain extent. After drug application, Compound B could significantly inhibit the increase of S100A8, S100A9, and K6 factors (P<0.05, P<0.01, P<0.001); compared with the TPA + positive drug group, the effect of 0.5% Compound B was comparable in inhibiting the expression of S100A8 and S100A9 factors, and the ear inflammation was alleviated.

[0203] 2.4.2.3 mRNA Expression of IL-1α, IL-1β, IL-4, IL-13, CXCL1 and CXCL2 in Mice with Irritant Dermatitis Treated with Matrix, Compound B (0.1%, 0.25%, 0.5%, 1% and 3%), and Positive Drug As Figure 59 shown, after TPA treatment, the mRNA expression levels of IL-1α, IL-1β, IL-4, IL-13 and CXCL2 in skin tissues increased, and Compound B could significantly inhibit the increase of the above genes to a certain extent (P<0.05, P<0.01, P<0.001, P<0.0001). Compared with the TPA+positive drug group, 0.5% Compound B had no obvious difference in reducing the expression of IL-1α, IL-1β, IL-4 and IL-13 factors, and had a local anti-inflammatory effect on ear inflammation.

[0204] 2.4.2.4 mRNA Expression of IL-17A, IL-22 and IL-23A in Mice with Irritant Dermatitis Treated with Matrix, Compound B (0.1%, 0.25%, 0.5%, 1% and 3%), and Positive Drug As Figure 60 shown, after TPA treatment, the mRNA expression levels of IL-17A, IL-22 and IL-23A in skin tissues increased, and Compound B could significantly inhibit the contents of IL-17A, IL-22 and IL-23A (P<0.05, P<0.01, P<0.001). Among them, 0.5% Compound B had the best effect on improving the immune function of the ear. 2.4.2.5 mRNA Expression of VEGF-A and ICAM-1 in Mice with Irritant Dermatitis Treated with Matrix, Compound B (0.1%, 0.25%, 0.5%, 1% and 3%), and Positive Drug As Figure 61 shown, after TPA treatment, the mRNA expression level of the vascular permeability-related factor VEGF-A increased, and 0.5% Compound B could significantly reduce the expression of VEGF-A in skin tissues (P<0.05).

[0205] 2.4.3 Application Effects of Different Concentrations of Compound C on a Mouse Model of Irritant Dermatitis 2.4.3.1 Effects of Matrix, Compound C (0.01%, 0.1%, 0.25%, 0.5%, 1%, 3% and 5%), and Positive Drug on the Ear Margin Thickness of Mice in Cases of Irritant Dermatitis As Figure 62As shown, after topical application of TPA to the ears of mice, morphological changes in the ears of mice were visible to the naked eye. Redness, thickening, and edema appeared in the ears of the TPA + matrix group. Through ear measurement, before the experiment, there were no differences in the thickness of the ear margins among the groups. After modeling, the thickness of the ear margins of all groups of mice increased to varying degrees, and the increase was significant in the TPA + matrix group, indicating the occurrence of inflammatory edema in the ears of mice. Compared with the TPA + matrix group, the thickness of the ear margins in each drug administration group decreased significantly (P < 0.05, P < 0.01, P < 0.001), and the inhibitory effect of 0.5% compound C was the most obvious, which could reduce the ear edema phenomenon.

[0206] Among them, two concentrations of 0.01% and 5% were used to screen the effective concentrations of the compound. The results showed that the effect of 0.01% was relatively poor compared with 0.1%, and the effect of 5% was relatively poor compared with 3%. Therefore, for the consideration of optimizing the experimental design and cost - effectiveness, the efficacy of the compound in the concentration range of 0.1% - 3% was mainly explored in the mouse model of irritant dermatitis subsequently.

[0207] 2.4.3.2 mRNA expression of S100A8, S100A9, and K6 in the mouse model of irritant dermatitis with matrix, compound C (0.1%, 0.25%, 0.5%, 1%, and 3%), and positive drug As Figure 63 shown, the mRNA of S100A8, S100A9, and K6 increased significantly after TPA treatment, indicating that TPA treatment damaged the epidermal barrier function, which led to the occurrence of ear inflammation to a certain extent. After application by the drug administration group, compound C could significantly inhibit the increase of S100A8, S100A9, and K6 factors (P < 0.05, P < 0.01, P < 0.001). Compared with the TPA + positive drug group, 0.5% compound C had an equivalent effect in inhibiting the expression of the S100A8 factor and alleviated ear inflammation.

[0208] 2.4.3.3 mRNA expression of IL - 1α, IL - 1β, IL - 4, IL - 13, CXCL1, and CXCL2 in the mouse model of irritant dermatitis with matrix, compound C (0.1%, 0.25%, 0.5%, 1%, and 3%), and positive drug As Figure 64 shown, after TPA treatment, the mRNA expression levels of IL - 1α, IL - 1β, IL - 4, IL - 13, and CXCL2 in the skin tissue increased, and compound C could inhibit the increase of these genes (P < 0.05, P < 0.01, P < 0.001). Compared with the TPA + positive drug group, 0.5% compound C had an equivalent effect in reducing the expression of IL - 1α and IL - 1β factors and had a local anti - inflammatory effect on ear inflammation.

[0209] 2.4.3.4 mRNA Expression of IL-17A, IL-22 and IL-23A in Mice with Irritant Dermatitis Treated with Matrix, Compound C (0.1%, 0.25%, 0.5%, 1% and 3%), and Positive Drug As Figure 65 shown, after TPA treatment, the mRNA expression levels of IL-17A, IL-22 and IL-23A in skin tissues increased. Compound C at 0.5% could inhibit the expression of IL-17A, IL-22 and IL-23A to a certain extent (P<0.05, P<0.01, P<0.001), and had an improving effect on the immune function of the ear. 2.4.3.5 mRNA Expression of VEGF-A and ICAM-1 in Mice with Irritant Dermatitis Treated with Matrix, Compound C (0.1%, 0.25%, 0.5%, 1% and 3%), and Positive Drug.

[0210] As Figure 66 shown, after TPA treatment, the mRNA expression level of the vascular permeability-related factor VEGF-A increased. Compound C had a tendency to reduce the expression of VEGF-A in skin tissues, and Compound C at 0.5% could significantly reduce the expression of VEGF-A (P<0.05).

[0211] Example 5 Application of Hesperidin Derivatives in Improving Rabbit Ear Scar Model 1. Experimental Materials 1.1 Experimental Animals New Zealand White Rabbits 1.2 Experimental Reagents Compound A Group (0.01% Compound A, 0.1% Compound A, 0.25% Compound A, 0.5% Compound A, 1% Compound A, 3% Compound A and 5% Compound A); Compound B Group (0.01% Compound B, 0.1% Compound B Group, 0.25% Compound B Group, 0.5% Compound B Group, 1% Compound B Group, 3% Compound B Group and 5% Compound B); Compound C (0.01% Compound C, 0.1% Compound C, 0.25% Compound C, 0.5% Compound C, 1% Compound C, 3% Compound C and 5% Compound C); Positive Control: Kelo-cote Gel.

[0212] 2. Experimental Methods 2.1 Establishment of Rabbit Ear Scar Model and Drug Administration Method Six New Zealand white rabbits of appropriate age were anesthetized intravenously. Along the long axis on the ventral side of each rabbit ear, avoiding blood vessels, four circular wounds with a diameter of about 6 mm were made with a circular drill bit, and the full-thickness skin was excised to reach the surface of the perichondrium. The distance between each wound was about 1 cm. After the operation, routine anti-infection measures were taken (800,000 U of penicillin was given by intramuscular injection in the buttocks on the same day to prevent infection), and the rabbits were allowed to eat and drink freely. After 30 days, the wounds healed and protruded significantly above the skin surface, forming hypertrophic scars.

[0213] 2.2 Observation indicators 2.2.1 Scar healing time: Observe the healing situation of the rabbit ear wounds, the wound healing time, the formation of hyperplastic masses, and the appearance rate of hyperplastic mass tissues in different parts (near, middle, and far) of the rabbit ear wounds within a certain period.

[0214] 2.2.2 Number of capillaries and fibroblasts in wound granulation tissue: On the 7th day and the 14th day respectively, tissue specimens were taken from the wound edges of the rabbit ears in each group and observed under an ordinary optical microscope. Five complete and non-repeating high-power microscopic fields were randomly selected from each section to count the number of capillaries and fibroblasts in the wound granulation tissue, and their mean values were used as the measurement values of the section. At the same time, the number of fibroblasts in the scar tissue of the rabbit ears was taken when observing the scars for 3 months.

[0215] 2.3 Statistical processing Same as Example 1 2.4 Research results 2.4.1 Application effects of compound A at different concentrations on the rabbit ear wound model 2.4.1.1 Scar healing time Since the healing times of different groups were different, the day of complete healing was taken as the first day of scar observation. Using three indicators: 1. The skin color was approximately the same as other parts of the body; 2. The maximum vertical distance of the scar above the normal skin surface was less than 1 mm; 3. The scar was soft. Observe for 3.5 months. If the scar was not completely treated after 3.5 months, it was recorded as 3.5 months. The scar healing time is shown in Table 2: Table 2 Scar healing time

[0216] ** Compared with the blank control group P <0.01; * Compared with the blank control group P <0.05 Based on the experimental data, the two concentrations of 0.01% and 5% were used to screen the effective concentrations of the compound. The results showed that the effect of 0.01% was relatively poor compared to 0.1%, and the effect of 5% was relatively poor compared to 3%. Therefore, considering the optimization of the experimental design and cost-effectiveness, the efficacy of the compound in the concentration range of 0.1% - 3% was mainly explored in the rabbit ear wound model in the follow-up.

[0217] 2.4.1.2 The number of capillaries and fibroblasts is shown in Tables 3 and 4 as follows: Table 3 The number of capillaries in wound granulation tissue

[0218] ** P < 0.01 compared with the blank control group; * P < 0.05 compared with the blank control group Table 4 The number of fibroblasts in wound granulation tissue

[0219] *** Compared with the blank control group P < 0.001; ** compared with the blank control group P < 0.01; * compared with the blank control group P < 0.05 The rapid growth of capillaries and fibroblasts indicates that the wound is in a period of rapid healing. However, the number of fibroblasts after wound healing also indicates the size of the wound scar, that is, the more excessive proliferation of fibroblasts, the larger the wound scar. In the observation on the 14th day, the number of fibroblasts in each drug administration group showed a relatively low level. As the concentration of Compound A increased, the drug effect gradually enhanced, but when the concentration exceeded 0.5%, the drug effect decreased instead. Among them, Compound A at a concentration of 0.5% showed the most significant drug effect. It shows that the product of the present invention not only has a significant effect on healing wounds, but also has a certain inhibitory effect on the formation of scars. In the blank control group, since the wound is still healing, the fibroblasts are in a growing state. For the 3-month observation of scar treatment, the more excessive proliferation of fibroblasts, the larger the wound scar.

[0220] 2.4.2 Application effects of different concentrations of Compound B on rabbit ear wound models The scar healing time is shown in Table 5, and the number of capillaries and fibroblasts in wound granulation tissue are shown in Tables 6 and 7 respectively.

[0221] Table 5 Scar healing time

[0222] Based on the experimental data, two concentrations of 0.01% and 5% were used to screen the effective concentration of the compound. The results showed that 0.01% had a relatively poor effect compared with 0.1%, and 5% had a relatively poor effect compared with 3%. Therefore, considering the optimization of the experimental design and cost-benefit, the compound efficacy in the concentration range of 0.1% - 3% was mainly explored in the rabbit ear wound model subsequently.

[0223] Table 6 The number of capillaries in wound granulation tissue

[0224] Table 7 The number of fibroblasts in wound granulation tissue

[0225] *** Compared with the blank control group P < 0.001; ** Compared with the blank control group P < 0.01; * Compared with the blank control group P < 0.05 In the data observation on the 14th day, the number of fibroblasts in all drug administration groups decreased. As the concentration of Compound B increased, the drug efficacy gradually enhanced, but when the concentration exceeded 0.5%, the drug efficacy decreased instead. Among them, Compound B at a concentration of 0.5% showed the most significant drug efficacy. Compound B at a concentration of 0.5% was slightly inferior to the positive control drug, BAKER'S SILICONE GEL, in terms of drug efficacy. This result indicates that the product of the present invention has a certain therapeutic effect in promoting wound healing and inhibiting scar formation. In the blank control group, since the wound was still healing, the fibroblasts were in a growing state. For the 3-month observation of scar treatment, the more excessive proliferation of fibroblasts, the larger the wound scar.

[0226] 2.4.3 Application effects of different concentrations of Compound C on the rabbit ear wound model The scar healing time is shown in Table 8, and the number of capillaries and fibroblasts in wound granulation tissue are shown in Table 9 and Table 10 respectively.

[0227] Table 8 Scar healing time

[0228] ** Compared with the blank control group P < 0.01; * Compared with the blank control group P < 0.05 Based on the experimental data, two concentrations of 0.01% and 5% were used to screen the effective concentration of the compound. The results showed that 0.01% was less effective than 0.1%, and 5% was less effective than 3%. Therefore, for the consideration of optimizing the experimental design and cost-effectiveness, the compound efficacy in the concentration range of 0.1% - 3% was mainly explored in the rabbit ear wound model subsequently.

[0229] Table 9 The number of capillaries in wound granulation tissue

[0230] Table 10 The number of fibroblasts in wound granulation tissue

[0231] *** Compared with the blank control group P < 0.001; ** Compared with the blank control groupP < 0.01; *Compared with the blank control group P < 0.05 In the data observation on the 14th day, the number of fibroblasts in all drug administration groups decreased. As the concentration of Compound C increased, the drug efficacy gradually enhanced, but when the concentration exceeded 0.5%, the drug efficacy decreased instead. Among them, Compound C at a concentration of 0.5% showed the most significant drug efficacy. Compound C at 0.5% was slightly inferior to the positive control drug, silicone gel, in terms of drug efficacy performance. This result indicates that the product of the present invention has a certain therapeutic effect in promoting wound healing and inhibiting scar formation. In the blank control group, since the wound was still healing, the fibroblasts were in a growing state. For the 3-month observation of scar treatment, the more excessive proliferation of fibroblasts, the larger the wound scar.

[0232] 2.5 Conclusion By comparing in detail the number of days required for scar healing, the growth of fibroblasts at the wound site, and the relevant indicators of capillary neovascularization, it is known that there are differences in the drug efficacy of different compounds in promoting wound healing. Specifically, Compound A at 0.5% exhibited the most excellent drug efficacy. It not only accelerated the scar healing time but also performed outstandingly in promoting the proliferation of fibroblasts at the wound and inducing capillary neovascularization, and the results far exceeded those of other test substances. In summary, the drug efficacy ranking from high to low is: Compound A > positive drug > Compound B ≈ Compound C The various embodiments of the present invention have been described above. The above description is exemplary and not exhaustive, and is not limited to the disclosed embodiments. Many modifications and variations are obvious to those of ordinary skill in the art in the technical field without departing from the scope and spirit of the described embodiments. The selection of the terms used herein is intended to best explain the principles of the embodiments, practical applications, or improvements to the technologies in the market, or to enable other ordinary skill in the art in the technical field to understand the embodiments disclosed herein.

Claims

1. Use of a hesperidin derivative in the preparation of a product for treating skin-lesion dermatosis, characterized in that, The hesperidin derivatives are selected from glucosyl hesperidin, methyl hesperidin chalcone or neohesperidin dihydrochalcone.

2. The application according to claim 1, characterized in that, The concentration of the hesperidin derivative in the product is 0.01% - 5%.

3. The application according to claim 1, characterized in that, The skin lesion skin disease is psoriasis.

4. The application according to claim 3, characterized in that, The concentration of the hesperidin derivative in the product is 0.1% - 2%.

5. The application according to claim 1, wherein The skin lesion skin disease is atopic dermatitis, irritant dermatitis, skin damage caused by sensitive skin or scars.

6. The application according to claim 5, wherein The concentration of the hesperidin derivative in the product is 0.1% - 3%.

7. The application according to claim 1, wherein The product is the hesperidin derivative, or the product comprises: the hesperidin derivative and a pharmaceutically or cosmetically acceptable carrier.

8. The application according to claim 1, characterized in that, The dosage form of the product includes: ointment, spray, liniment, patch, emulsion, gel or lotion.

9. The application according to claim 1, characterized in that, The product exerts a therapeutic effect through at least one of the following mechanisms: Regulating the expression levels of S100A8, S100A9, and K6 mRNA; Reducing the expression levels of IL-1α, IL-1β, CXCL1, and CXCL2; Reducing the expression levels of IL-17A, IL-22, and IL-23A; Reducing the expression levels of VEGF-A and ICAM-1.

10. The application according to claim 1, characterized in that, The product further contains at least one active ingredient selected from glucocorticoids, calcineurin inhibitors, antibiotics, antihistamines, and immunosuppressants.

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