Use of cembranoid diols in the preparation of a medicament for the prevention and treatment of rosacea

Abstract

The application relates to application of cedrene diol in preparation of medicines for preventing and treating rosacea, and belongs to the technical field of biological medicines. The application provides application of cedrene diol in preparation of medicines for preventing and / or treating rosacea. The cedrene diol can significantly improve redness and inflammation of an injection area of a rosacea mouse model in long and short terms, can significantly relieve skin histopathological inflammation of the rosacea mouse model (including improving thickening of skin epidermis of the rosacea mouse model, reducing inflammatory cell infiltration of skin tissue of the rosacea mouse model, and reducing capillary proliferation of skin tissue of the rosacea mouse model), can significantly reduce skin inflammation of a rosacea patient (including reducing pain of the skin of the rosacea patient and relieving redness of the skin of the rosacea patient), and can significantly improve skin damage of the rosacea patient (including improving papules and pustules of the skin of the rosacea patient).

Description

Technical Field

[0001] This invention relates to the application of cepertrienol in the preparation of drugs for the prevention and treatment of rosacea, and belongs to the field of biomedical technology. Background Technology

[0002] Rosacea, also known as rhinophyma, is a chronic inflammatory skin disease that commonly affects the central face, primarily involving facial blood vessels and the peri-pillar sebaceous units. Clinical manifestations mainly include skin flushing, erythema, papules, pustules, and telangiectasia. Rosacea is most common in middle-aged individuals between 30 and 50 years old, with women being more frequently affected than men, although severe cases are generally seen in men. The causes of rosacea are complex, and various factors can trigger or worsen the disease, including neurovascular dysfunction, impaired skin barrier function, immune inflammatory responses, and endocrine disorders. Unhealthy or irregular diets, as well as high stress levels and psychological factors, can also trigger rosacea. The course of rosacea is slow, typically divided into three stages: the erythematous telangiectatic stage, the papulopustular stage, and the rhinophyma stage, with no clear boundaries between the stages. Patients in the erythematous telangiectatic and papulopustular stages can recover with medication, while patients in the rhinophyma stage require additional surgical intervention such as excision or dermabrasion to restore their appearance. Therefore, timely and effective drug intervention for rosacea is still necessary to prevent it from progressing to the rhinophyma stage.

[0003] Currently, rosacea is mainly treated with antibiotics (including doxycycline and minocycline), hydroxychloroquine, isotretinoin, beta-adrenergic receptor blockers (including carvedilol and metoprolol), endocrine preparations (including diethylstilbestrol and birth control pills), and anti-anxiety drugs (including paroxetine and alprazolam). However, these drugs still have some drawbacks. For example, antibiotics, hydroxychloroquine, beta-adrenergic receptor blockers, endocrine preparations, and anti-anxiety drugs all have significant side effects. While isotretinoin has fewer side effects, it requires long-term use, and its effect on relieving rosacea is not ideal, only playing a certain auxiliary role. Therefore, there is an urgent need to find more rosacea treatments with fewer side effects that can effectively relieve rosacea symptoms (including relieving skin erythema caused by rosacea, relieving skin inflammation caused by rosacea, and improving skin damage caused by rosacea).

[0004] Tobacco, the leaf of the Nicotiana tabacum L. (a plant in the Solanaceae family), has the effects of regulating qi and relieving pain, as well as drying dampness (referring to the traditional Chinese medicine method of removing dampness with drying herbs). It is often used to treat qi stagnation pain, joint pain, carbuncles, and scabies. β-Cembrenediol (β-CBD, CAS: 57605-81-9) is an active substance extracted from tobacco. Studies have shown that β-Cembrenediol not only does not pose a threat to human health but also possesses various biological activities such as anti-tumor, neuroprotective, antibacterial, and insecticidal effects, exhibiting high safety. Furthermore, β-Cembrenediol is readily available and inexpensive. Currently, there are no reports on the use of β-Cembrenediol for the prevention and / or treatment of rosacea. Summary of the Invention

[0005] To address the above problems, this invention provides the use of cepertrienol in the preparation of medicaments for the prevention and / or treatment of rosacea.

[0006] In one embodiment of the present invention, the prevention / or treatment of rosacea includes improving skin erythema caused by rosacea, improving skin inflammation caused by rosacea, and / or improving skin damage caused by rosacea.

[0007] In one embodiment of the present invention, the improvement of skin inflammation caused by rosacea includes relieving epidermal thickening caused by rosacea, improving inflammatory cell infiltration in skin tissue caused by rosacea, improving capillary hyperplasia caused by rosacea, relieving skin pain caused by rosacea, and / or relieving skin redness and swelling caused by rosacea.

[0008] In one embodiment of the present invention, the improvement of skin lesions caused by rosacea includes improving acne, papules, pustules, nodules, cysts and / or scars caused by rosacea.

[0009] In one embodiment of the present invention, the drug comprises an active ingredient and pharmaceutically acceptable excipients; the active ingredient comprises cepertrienildiol.

[0010] In one embodiment of the present invention, the active component is cepertrienildiol.

[0011] In one embodiment of the present invention, the pharmaceutically acceptable excipients include solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, binding agents, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculants, anti-flocculation agents, filter aids, and / or release inhibitors.

[0012] In one embodiment of the present invention, the dosage form of the drug is a liniment; the liniment includes bath preparations, lotions, liniments, tinctures, oils, plasters, ointments, pastes, powders, coatings, films, gels, and / or latex patches.

[0013] The present invention also provides a medicament for the prevention and / or treatment of rosacea, the medicament containing an active ingredient; the active ingredient comprising cepertrienildiol.

[0014] In one embodiment of the present invention, the prevention / or treatment of rosacea includes improving skin erythema caused by rosacea, improving skin inflammation caused by rosacea, and / or improving skin damage caused by rosacea.

[0015] In one embodiment of the present invention, the improvement of skin inflammation caused by rosacea includes relieving epidermal thickening caused by rosacea, improving inflammatory cell infiltration in skin tissue caused by rosacea, improving capillary hyperplasia caused by rosacea, relieving skin pain caused by rosacea, and / or relieving skin redness and swelling caused by rosacea.

[0016] In one embodiment of the present invention, the improvement of skin lesions caused by rosacea includes improving acne, papules, pustules, nodules, cysts and / or scars caused by rosacea.

[0017] In one embodiment of the present invention, the active component is cepertrienildiol.

[0018] In one embodiment of the invention, the drug further comprises pharmaceutically acceptable excipients.

[0019] In one embodiment of the present invention, the pharmaceutically acceptable excipients include solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, binding agents, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculants, anti-flocculation agents, filter aids, and / or release inhibitors.

[0020] In one embodiment of the present invention, the dosage form of the drug is a liniment; the liniment includes bath preparations, lotions, liniments, tinctures, oils, plasters, ointments, pastes, powders, coatings, films, gels, and / or latex patches.

[0021] The technical solution of this invention has the following advantages:

[0022] This invention provides the application of cephalotrimazole in the preparation of drugs for the prevention and / or treatment of rosacea. Studies have found that cephalotrimazole can significantly improve erythema and inflammation at the injection site in mouse models of rosacea in both long-term and short-term conditions; significantly alleviate skin histopathological inflammation in mouse models of rosacea (including improving epidermal thickening, reducing inflammatory cell infiltration, and reducing capillary proliferation); significantly reduce skin inflammation in patients with rosacea (including reducing pain and redness); and significantly improve skin lesions in patients with rosacea (including improving papules and pustules). Therefore, cephalotrimazole shows great promise in the preparation of drugs for the prevention and / or treatment of rosacea. Attached Figure Description

[0023] Figure 1 Typical manifestations of the skin on the back of mice in different groups (short-term treatment manifestations).

[0024] Figure 2 Erythema scores (short-term treatment performance) at the injection site on the back of mice in different groups. Figure 2 In the mean, *P<0.01.

[0025] Figure 3 : Erythema area in the dorsal injection area of ​​mice in different groups (short-term treatment effect). Figure 3 In the mean square, *P<0.01.

[0026] Figure 4 Skin tissues from different groups of mice (H&E staining for pathological examination).

[0027] Figure 5 Inflammatory cell infiltration in the skin tissues of mice in different groups. Figure 5 In the mean square, *P<0.01.

[0028] Figure 6 Skin tissues from different groups of mice (observed and examined for pathological results by toluidine blue staining).

[0029] Figure 7 Mast cell infiltration in the skin tissues of mice in different groups. Figure 7 In the mean square, *P<0.01.

[0030] Figure 8 Typical manifestations of the skin on the back of mice in different groups (long-term treatment manifestations).

[0031] Figure 9 Erythema scores (long-term treatment performance) at the injection site on the back of mice in different groups. Figure 9 In the mean square, *P<0.01.

[0032] Figure 10 : Erythema area in the dorsal injection area of ​​mice in different groups (long-term treatment performance). Figure 10 In the mean square, *P<0.01.

[0033] Figure 11 Changes in skin lesions in rosacea patients after different durations of medication use. Detailed Implementation

[0034] The following embodiments are provided to better understand the present invention and are not limited to the preferred embodiments described. They do not constitute a limitation on the content and scope of protection of the present invention. Any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention.

[0035] For any experimental steps or conditions not specified in the following examples, the procedures or conditions described in the literature in this field can be followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional reagent products.

[0036] Experimental Example 1: Effects of cepertrienol on a mouse model of rosacea (short-term treatment performance)

[0037] The experimental procedure is as follows:

[0038] Male BALB / c mice (7 weeks old, 18–22 g) used in the experiment were purchased from Beijing Vital River Company. Hulling conditions: constant temperature 24±2℃, humidity 55%±15%, 12-hour light / dark cycle, free access to water and food. All animal use was approved by the International Association for Laboratory Animal Management and Use (IACUC). After one week of acclimatization in the laboratory, the mice underwent hair removal (2×2 cm) on their backs using a combination of electric shaver and depilatory cream. Experiments began 24 hours after hair removal.

[0039] The experiment was divided into two groups: a PBS control group and a β-CBD treatment group, with 5 mice in each group. After grouping, the mice were operated on. In the PBS control group, 40 μL of LL-37 solution (LL-37 dry powder was prepared into a 320 μmol / L solution with PBS buffer; LL-37 dry powder was purchased from MedChemExpress, catalog number: HY-P5391) was injected intradermally into the hairless area on the mouse's back, penetrating 6 mm along the skin. This injection was recorded as the first injection (to induce a rosacea-like erythema phenotype in mice through the antimicrobial peptide LL-37). Two hours after the first injection, 50 μL of PBS buffer (PBS buffer was purchased from Therm) was applied to the injection area on the mouse's back. oFisher (product number: 10010023) was used, and this application was recorded as the first intervention. Twelve hours after the first injection, a second injection was given, with the same dosage and method as the first injection. Two hours after the second injection, a second intervention was given, with the same dosage and method as the first intervention. Twelve hours after the second injection, a third injection was given, with the same dosage and method as the second injection. Two hours after the third injection, a third intervention was given, with the same dosage and method as the second intervention. Twelve hours after the third injection, a fourth injection was given, with the same dosage and method as the third injection. Two hours after the fourth injection, a fourth intervention was given, with the same dosage and method as the third intervention.

[0040] The procedure for the β-CBD treatment group was as follows: 40 μL of LL-37 solution (LL-37 dry powder was prepared to a 320 μmol / L solution with PBS buffer) was drawn into an insulin needle and injected intradermally into the hairless area on the mouse's back, penetrating 6 mm along the skin. This injection was recorded as the first injection (to induce a rosacea-like erythema phenotype in mice via the antimicrobial peptide LL-37). Two hours after the first injection, 50 μL of cipertrienol solution (cipertrienol was prepared to a 1 mg / mL solution with PBS buffer; cipertrienol was purchased from Cayman, catalog number: 57) was applied to the injection area on the mouse's back. (605-81-9), this application was recorded as the first intervention; 12 hours after the first injection, a second injection was given, with the same dosage and method as the first injection; 2 hours after the second injection, a second intervention was given, with the same dosage and method as the first intervention; 12 hours after the second injection, a third injection was given, with the same dosage and method as the second injection; 2 hours after the third injection, a third intervention was given, with the same dosage and method as the second intervention; 12 hours after the third injection, a fourth injection was given, with the same dosage and method as the third injection; 2 hours after the fourth injection, a fourth intervention was given, with the same dosage and method as the third intervention.

[0041] Twelve hours after the fourth injection, the mice were euthanized by cervical dislocation, and photographs were taken of the skin on their backs. The results are shown below. Figure 1 The severity of erythema in mice was scored, and the scores are shown below. Figure 2 The area of ​​erythema on mouse skin lesions was counted, and the results are shown in [see attached table]. Figure 3 The severity of erythema was scored from 0 to 4 points, with 0 indicating no erythema, 1 indicating faintly visible erythema, 2 indicating pale erythema, 3 indicating obvious and clear erythema, and 4 indicating deep erythema with clear borders. Mouse skin was fixed in formalin, embedded in paraffin, sectioned (4 μm thick), stained with hematoxylin and eosin (H&E), and subjected to histopathological examination. The results are shown in the table below. Figure 4 Five fields of view were randomly selected from the HE-stained sections under a 400x microscope. The number of cells in each field of view was counted using ImageJ software, and the average value was taken. The results are shown below. Figure 5 Mouse skin was fixed in formalin, embedded in paraffin, sectioned (4 μm thick), stained with toluidine blue, and subjected to histopathological examination. The results are shown below. Figure 6 (Mast cells are a type of inflammatory cell. Among inflammatory cells, rosacea is more closely related to mast cells. Therefore, toluidine blue staining was used here to further observe changes in mast cells and thus assess the effect of drugs on skin inflammation caused by rosacea.) Five fields of view were randomly selected under a 400x microscope after toluidine blue staining. ImageJ software was used to count the number of cells in each field of view, and the average value was taken. The statistical results are shown below. Figure 7 .

[0042] Figure 1 The experimental results showed that after continuous administration of LL-37, obvious erythema phenotype appeared on the skin of the injection area on the back of mice. After intervention with cepertrienil, the area of ​​erythema on the skin of the injection area on the back of mice was significantly reduced. Figure 2 The experimental results showed that the erythema score of the dorsal injection area of ​​mice in the PBS group was 3.4±0.55, while that in the β-CBD group was 1.4±0.55. The erythema score of the dorsal injection area of ​​mice in the PBS group was significantly higher than that in the β-CBD group (P<0.01). Figure 3 The experimental results showed that the erythema area in the dorsal injection area of ​​mice in the PBS group was 0.34 ± 0.069 cm². 2 The erythema area in the dorsal injection area of ​​mice in the β-CBD group was 0.09 ± 0.023 cm². 2 The area of ​​erythema in the dorsal injection area of ​​mice in the PBS group was significantly higher than that in the β-CBD group (P<0.01).

[0043] Figure 4 The experimental results showed that the skin epidermis of mice in the PBS group was significantly thickened, with a large number of inflammatory cell infiltrations and obvious capillary proliferation (indicated by the arrow). Compared with the PBS group, the skin epidermis of mice in the β-CBD group was thinner, with reduced inflammatory cell infiltrations and no obvious capillary proliferation. Figure 5 The experimental results showed that the number of inflammatory cells infiltrating the skin of mice in the PBS group (381.4±67.4 / field of view) was significantly higher than that in the β-CBD group (80.4±11.26 / field of view) (P<0.01). Figure 6 The experimental results showed that mast cell infiltration in the skin tissue of mice in the PBS group was significantly increased (cells indicated by arrows), while mast cell infiltration in the skin tissue of mice in the β-CBD group was significantly reduced compared with the PBS group. Figure 7 The experimental results showed that the number of mast cells infiltrating the skin of mice in the PBS group (16.4±1.67 / field of view) was significantly higher than that in the β-CBD group (5.6±0.89 / field of view) (P<0.01).

[0044] comprehensive Figures 1 to 7 The experimental results show that cepertrienol can reduce erythema and inflammation in the injection area of ​​a mouse model of rosacea in the short term, and that cepertrienol can alleviate the pathological inflammation of the skin tissue in a mouse model of rosacea.

[0045] Experimental Example 2: Effects of cipertrienol on a mouse model of rosacea (long-term treatment performance)

[0046] The experimental procedure is as follows:

[0047] Male BALB / c mice (7 weeks old, 18–22 g) used in the experiment were purchased from Beijing Vital River Pharmaceutical Co., Ltd. Husbandry conditions: constant temperature 24±2℃, humidity 55%±15%, 12-hour light / dark cycle, free access to water and food. All animals used were approved by the International Association for Laboratory Animal Management and Use (IACUC). After acclimatization in the laboratory for one week, the mice were shaved (2×2 cm) from their backs using a combination of electric shaver and depilatory cream. Twenty-four hours after depilation, 40 μL of LL-37 solution (LL-37 dry powder prepared to a 320 μmol / L solution with PBS buffer) was injected intradermally into the shaved area on the mouse's back, penetrating 6 mm along the skin. Injections were given every 12 hours for a total of four injections (to induce a rosacea-like erythema phenotype in the mouse skin via the antimicrobial peptide LL-37). The experiment began 12 hours after the fourth injection.

[0048] The experiment was divided into two groups: a PBS control group and a β-CBD treatment group, with five mice in each group. After grouping, the mice were treated. In the PBS control group, 50 μL of PBS buffer was applied to the injection site on the back of the mice for intervention, twice daily for six consecutive days. In the β-CBD treatment group, 50 μL of cipertrienol solution (cipertrienol prepared to a 1 mg / mL solution with PBS buffer) was applied to the injection site on the back of the mice for intervention, twice daily for six consecutive days.

[0049] During the intervention period, the skin on the backs of the mice was photographed daily, and the results are shown below. Figure 8 After the intervention, the severity of erythema in the mice was assessed, and the scores are shown below. Figure 9 The area of ​​erythema on mouse skin lesions was counted, and the results are shown in [see attached table]. Figure 10 The severity of erythema is scored from 0 to 4 points, with 0 indicating no erythema, 1 indicating faintly visible erythema, 2 indicating light erythema, 3 indicating obvious and clear erythema, and 4 indicating deep erythema with clear borders.

[0050] Figure 8 The experimental results showed that on the fourth day after intervention, there were differences in erythema and inflammation in the injection area on the back of the two groups of mice. The PBS group mice showed obvious erythema phenotype in the injection area on the back, while the erythema area in the injection area on the back of the β-CBD group mice was significantly reduced. On the sixth day after intervention, the differences in erythema and inflammation in the injection area on the back of the two groups of mice were more obvious. The erythema in the injection area on the back of the PBS group mice was still very obvious, while the erythema in the injection area on the back of the β-CBD group mice almost disappeared completely.

[0051] Figure 9The experimental results showed that on the fourth day after intervention, there was a difference in the erythema scores of the dorsal injection areas of the two groups of mice. The erythema scores of the dorsal injection areas of the PBS group mice continued to increase compared with the previous three days, while the erythema scores of the dorsal injection areas of the β-CBD group mice decreased significantly compared with the previous three days. On the sixth day after intervention, the difference in erythema scores of the dorsal injection areas of the two groups of mice was more obvious. The erythema scores of the dorsal injection areas of the PBS group mice were still 3 points, while the erythema scores of the dorsal injection areas of the β-CBD group mice decreased to 1 point. The erythema scores of the dorsal injection areas of the PBS group mice were significantly higher than those of the β-CBD group (P<0.01).

[0052] Figure 10 The experimental results showed that there were significant differences in the change of erythema area in the dorsal injection area between the two groups of mice during the intervention period. In the PBS group, the erythema area in the dorsal injection area first increased slowly during the first four days of intervention, then began to decrease slowly on the fifth day, and finally decreased to 0.12 cm on the sixth day of intervention. 2 In the β-CBD group, the erythema area in the dorsal injection area of ​​mice continued to decrease over the six days of intervention, with the rate of decrease significantly increasing on the fifth day and finally decreasing to 0.04 cm on the sixth day. 2 The area of ​​erythema in the dorsal injection area of ​​mice in the PBS group was significantly higher than that in the β-CBD group (P<0.01).

[0053] comprehensive Figures 8-10 The experimental results show that cepertrienol can reduce erythema and inflammation in the injection area of ​​a mouse model of rosacea in the long term.

[0054] Experimental Example 3: Effects of cepertrienol on patients with rosacea

[0055] The experimental procedure is as follows:

[0056] The experiment recruited 10 volunteers with rosacea. The following is a typical case:

[0057] A 46-year-old male patient presented with a 5-year history of rosacea, primarily characterized by recurrent facial flushing, papules, and pustules. Topical application of cipertrienol solution (prepared with PBS buffer, concentration 50 μg / mL) to the affected areas was administered twice daily (morning and evening) for 15 consecutive days. After treatment, the patient reported significant pain relief, a marked reduction in papules and pustules, and improved flushing. No recurrence of symptoms occurred within six months. Figure 11 Similar to this patient, the symptoms of the other nine rosacea volunteers also improved significantly.

[0058] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. The use of cepertrienol in the preparation of medicaments for the prevention and / or treatment of rosacea, characterized in that, The prevention / or treatment of rosacea includes improving skin erythema caused by rosacea, improving skin inflammation caused by rosacea, and / or improving skin lesions caused by rosacea.

2. The application as described in claim 1, characterized in that, The drug comprises an active ingredient and pharmaceutically acceptable excipients; the active ingredient comprises cepertrienidol.

3. The application as described in claim 2, characterized in that, Pharmaceutically acceptable excipients include solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, binding agents, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculants, anti-flocculation agents, filter aids, and / or release inhibitors.

4. The application as described in any one of claims 1 to 3, characterized in that, The dosage form of the drug is a transdermal drug delivery preparation; the transdermal drug delivery preparation includes bath preparations, lotions, liniments, tinctures, oils, plasters, ointments, pastes, powders, coatings, films, gels and / or latex patches.

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