Triamterene and compositions thereof for treatment and / or prevention of inflammatory skin diseases

Abstract

The invention provides application of a compound of triamterene or pharmaceutically acceptable salt thereof in preparation of a medicine for treating and / or preventing inflammatory skin diseases. Compared with the prior art, the triamterene, the compound of the triamterene or the composition containing the triamterene can be used for treating and / or preventing inflammatory skin diseases. The applicant finds that the triamterene can inhibit abnormal proliferation of skin keratinocytes and skin inflammation response for the first time, and the symptom of psoriasis is remarkably relieved. The research on an imiquimod-induced mouse psoriasis-like model finds that the triamtedin can effectively improve the symptoms such as epidermis thickening, inflammation infiltration and erythema scale of mouse skin lesion tissues. Besides, the triamterene and the Infliximab are combined for treating the psoriasis, and compared with independent use of the triamterene and the Infliximab, the treatment effect can be remarkably enhanced. Therefore, the triamterene can be used as an effective component of the medicine to be prepared into a medicinal preparation for treating the psoriasis.

Description

Technical Field

[0001] The present application relates to the field of compound pharmaceutical technology, and in particular to the use of a compound of triamterene or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating and / or preventing inflammatory skin diseases. Background Art

[0002] Psoriasis is a common chronic, recurrent immune-mediated skin disease characterized by scaly erythema of the skin, which may be accompanied by arthritis in severe cases. Its pathogenesis involves abnormal regulation of the immune system, especially overactivation of T cells, which leads to excessive proliferation and inflammation of skin cells. Genetic and environmental factors (such as infection, stress, trauma, etc.) also increase the risk of psoriasis.

[0003] Types of psoriasis include vulgaris, pustular, psoriatic arthritis and erythroderma, and the treatment methods are diverse, covering topical drugs (steroids, vitamin D analogs), phototherapy, systemic therapy (methotrexate, cyclosporine) and biologics (TNF-α, IL-17, IL-23 inhibitors, etc.). Among them, biologics provide an important treatment option for patients with moderate to severe disease. However, existing therapies have limitations. Topical treatment and phototherapy have limited effects on severe patients, and phototherapy has long-term safety issues; systemic drugs have good efficacy but serious side effects; although biologics are effective and accurate, they are expensive, some patients will develop drug resistance or failure, and are accompanied by increased risks of infection. Therefore, although these therapies are effective for most patients, new treatment options still need to be developed to overcome the shortcomings of existing treatments.

[0004] Ademine (ADE) is a diuretic commonly used to treat hypertension and edema. It is a sodium channel blocker that can inhibit the reabsorption of sodium ions in the distal convoluted tubule and collecting duct, thereby increasing the excretion of sodium and water in the urine. Recent studies have shown that ademine has certain potential effects in cardiovascular disease, anti-fibrosis, and anti-inflammatory. However, although there have been many studies on the pharmacological effects of ademine in these fields, there have been no reports on related studies as a drug for the treatment of immune diseases, especially skin inflammatory diseases such as psoriasis. Summary of the invention

[0005] In view of the above-mentioned technical limitations, the present application proposes the use of a compound of triamterene or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating and / or preventing inflammatory skin diseases; it overcomes the deficiencies and defects mentioned in the background technology.

[0006] The chemical structural formula of triamterene is shown in formula (I).

[0007]

[0008] To achieve the above objectives, this application adopts the following technical solutions:

[0009] The invention of the present application is to provide the use of a compound of triamterene or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating and / or preventing inflammatory skin diseases.

[0010] Optionally, in the above-mentioned use, the compound is one or more of the nicotinate of triamterene, triamterene dihydrate, and furosemide-triamterene salt.

[0011] Optionally, in the above use, the inflammatory skin disease is selected from psoriasis, acne, seborrheic dermatitis, pityriasis rosea, cutaneous small vessel vasculitis, perioral dermatitis, tinea capitis, tinea pedis and tinea corporis.

[0012] Optionally, in the above-mentioned use, the inflammatory skin disease is psoriasis.

[0013] Optionally, in the above-mentioned use, the preparation type of the drug is selected from any one of tablets, capsules, granules, micropills, pellets, oral liquids, soft capsules, water injections, powder injections, and infusions.

[0014] The unit dose of tablets, capsules, granules, micropills, pellets, oral liquids, and soft capsules is 25-250 mg; the unit dose of aqueous injections, powder injections, and infusions is 5-100 mg.

[0015] The second inventive point of the present application is to provide the use of a combination comprising one or more compounds selected from the following in the preparation of a medicament for treating and / or preventing inflammatory skin diseases: a compound of triamterene or a pharmaceutically acceptable salt thereof, a TNF antagonist.

[0016] Optionally, in the above-mentioned use, the inflammatory skin disease is psoriasis.

[0017] Optionally, in the above-mentioned use, the TNF antagonist is Infliximab.

[0018] Optionally, in the above use, the dosage of triamterene is 1-30 mg / kg, preferably 10 mg / kg; the dosage of infliximab is 10 mg / kg.

[0019] Optionally, in the above-mentioned use, the preparation type of the drug is selected from any one of tablets, capsules, granules, micropills, pellets, oral liquids, soft capsules, water injections, powder injections, infusions, ointments, gels, and microemulsions.

[0020] Compared with the prior art, this application has the following advantages:

[0021] The present application discloses the use of a compound of triamterene or a pharmaceutically acceptable salt thereof in the preparation of a drug for the treatment and / or prevention of inflammatory skin diseases. The applicant discovered for the first time that triamterene can inhibit the abnormal proliferation of skin keratinocytes and significantly relieve the symptoms of psoriasis. Through the study of the psoriasis-like model of mice induced by imiquimod, it was found that triamterene can effectively improve the symptoms such as epidermal thickening, inflammatory infiltration and erythematous scaling in the skin lesions of mice. In addition, the combination of triamterene and infliximab for the treatment of psoriasis can significantly enhance the therapeutic effect compared with the use of either alone. It shows that triamterene can be used as an active ingredient of the drug to prepare a pharmaceutical preparation for the treatment of psoriasis. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It was shown that triamterene (ADE) had an improving effect on imiquimod (IMQ)-induced psoriasis-like inflammatory lesions in mice; Figure 1 A is a representative photo of the skin lesion and the results of H&E staining. Figure 1 B is the thickness of the epidermis, Figure 1 C is the percentage of body weight, Figure 1 D is the clinical score for skin tissue damage such as skin scaling, nodules, and erythema.

[0023] Figure 2 It was shown that triamterene (ADE) had an inhibitory effect on the abnormal proliferation of keratinocytes in the imiquimod (IMQ)-induced psoriasis-like skin lesion model in mice, and had an inhibitory effect on the activation of the NF-κB inflammatory pathway.

[0024] Figure 3 The results show that triamterene (ADE) was tested for cytotoxicity on keratinocytes (HaCaT) and no significant toxicity was observed within the dose range.

[0025] Figure 4 It was shown that the combination of triamterene (ADE) and infliximab has an improving effect on imiquimod (IMQ)-induced psoriasis-like inflammatory lesions in mice; Figure 4 A is a representative photo of the skin lesion. Figure 4 B is the percentage of body weight, Figure 4 C is the clinical score for skin tissue damage such as skin scaling, nodules, and erythema. DETAILED DESCRIPTION

[0026] In order to make the purpose, technical solution and advantages of the present application more clear, the present application is further described in detail below. However, it should be understood that the description here is only used to explain the present application and is not used to limit the scope of the present application.

[0027] Unless otherwise defined, all technical terms and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present application, and the terms used herein in the specification of the present application are only for the purpose of describing specific embodiments and are not intended to limit the present application. The reagents and instruments used herein are all commercially available, and the characterization means involved can refer to the relevant descriptions in the prior art, which will not be repeated herein.

[0028] In order to further understand the present application, the present application is further described in detail below in conjunction with the best embodiment.

[0029] Example 1

[0030] Use of triamterene in preparing a medicament for treating and / or preventing psoriasis, with a dosage of 1 mg / kg.

[0031] Use of triamterene and infliximab in combination in the preparation of a medicament for treating and / or preventing psoriasis, wherein the dosage of triamterene is 1 mg / kg and the dosage of infliximab is 10 mg / kg.

[0032] Example 2

[0033] Use of triamterene in preparing a medicament for treating and / or preventing psoriasis, wherein the dosage is 30 mg / kg.

[0034] Use of triamterene and infliximab in combination in the preparation of a medicament for treating and / or preventing psoriasis, wherein the dosage of triamterene is 30 mg / kg and the dosage of infliximab is 10 mg / kg.

[0035] Example 3

[0036] Use of triamterene in preparing a medicament for treating and / or preventing psoriasis, wherein the dosage is 10 mg / kg.

[0037] Use of triamterene and infliximab in combination in the preparation of a medicament for treating and / or preventing psoriasis, wherein the dosage of triamterene is 10 mg / kg and the dosage of infliximab is 10 mg / kg.

[0038] The effect verification experiments of Examples 4-6 were all conducted based on the dosage used in Example 3.

[0039] Example 4

[0040] Intraperitoneal injection of triamterene improves imiquimod-induced psoriasis-like inflammatory lesions in mice:

[0041] 1. Experimental animals:

[0042] C57 / B6J female mice, 68 weeks, 18-22g.

[0043] 2. Experimental methods:

[0044] Take a normal 68-week-old C57 / B6J female mouse and inject 200300μl of anesthetic intraperitoneally according to body weight to anesthetize it. Shave the hair on the back of the mouse to form an exposure area of ​​about 2cm×2cm. Randomly and evenly group them according to body weight. Shaving is performed 12 days before the formal experiment. Weigh the weight of the mouse regularly every day, observe local skin changes, take photos and record them, and give a score of 03 according to the psoriasis area and disease severity (PASI) scoring standard, and the first day is recorded as 0 points. Finally, apply imiquimod cream. Except for the normal control group, other groups apply 5% imiquimod cream 62.5mg / mouse (where the actual content of imiquimod is 3.125mg) on ​​the shaved skin, once a day, for 4 consecutive days. And give corresponding treatment.

[0045] Group 1: normal group, blank control, without any treatment;

[0046] Group 2: Model group, given imiquimod;

[0047] Group 3: The drug-treated group was given imiquimod and 10 mg / kg triamterene was intraperitoneally injected.

[0048] The normal group and the model group were given an equal amount of PBS, and the drug group was given an intraperitoneal injection according to the dose, once a day, 200 μL each time.

[0049] After 4 days, the skin of the mouse back was taken and photographed to record the condition. The skin lesion tissue was removed and the subcutaneous fat tissue was removed, and fixed in 4% paraformaldehyde solution for the preparation of paraffin sections.

[0050] 3. Evaluation method:

[0051] The following mouse PASI criteria were established with reference to the PASI (Psoriasis area and severity index) standard to evaluate clinical severity: erythema; infiltration; scaling. Each feature was evaluated with 0.3 points: 0 = none; 1 = mild; 2 = moderate; 3 = severe. Each site was scored according to the following three clinical characteristics of skin lesions: erythema (E): red or dark red inflammatory plaques that fade when pressed. 0-no erythema is visible; 1-light red; 2-red; 3-very deep red. Infiltration (I): the tendency of the lesion to spread to the surrounding areas, with unclear boundaries and a sense of substance when pressed. 0-the lesion is flush with the normal skin; 1-the lesion is slightly higher than the surface of the normal skin; 2-moderate elevation, the edge of the plaque is round or sloped; 3-the lesion is highly thickened and the elevation is very obvious. Epidermal desquamation / scaling (desquamation, D): refers to the shedding of epidermal cells in pieces. 0 - no visible scales on the surface; 1 - some lesions are covered with scales, mainly fine scales; 2 - most lesions are completely or incompletely covered with scales, and the scales are flaky; 3 - all lesions are covered with scales, and the scales are very thick and layered. Exudation, dryness and itching are not included in the total score.

[0052] 4. Analysis of pharmacological experimental results:

[0053] like Figure 1 The results showed that intraperitoneal injection of 10 mg / kg of triamterene can significantly improve imiquimod-induced psoriasis-like inflammatory lesions and IMQ-induced hyperproliferation of epidermal keratinocytes without obvious toxic side effects, demonstrating certain effectiveness and safety.

[0054] like Figure 2 The immunohistochemical results shown in the figure indicate that intraperitoneal injection of 10 mg / kg of triamterene can significantly reduce the expression of proliferation-related proteins PCNA and KI-67 and inflammatory response-related protein p-p65.

[0055] Example 5

[0056] Triamterene has no significant toxicity to keratinocytes:

[0057] 1. Experimental cells:

[0058] HaCaT cells (human immortalized keratinocytes).

[0059] 2. Experimental methods:

[0060] HaCaT was inoculated into a 96-well plate, and DMEM complete medium containing 10% serum was added and placed in a CO2 constant temperature culture. After adhesion, it was replaced with complete medium containing 0.1, 0.3, 1, 3, 10, 30, and 100 μM triamterene and continued to be cultured for 24 hours. 10 μl MTT solution (5 mg / ml, i.e. 0.5% MTT) was added to each well and cultured for 4 hours. After terminating the culture, the culture medium in the well was carefully aspirated. 100 μl dimethyl sulfoxide was added to each well, and the plate was placed on a shaker and oscillated at a low speed for 10 minutes to fully dissolve the crystals. The absorbance of each well was measured at OD490nm on an enzyme-linked immunosorbent assay. At the same time, zero wells (culture medium, MTT, dimethyl sulfoxide) and control wells (cells, drug dissolution medium of the same concentration, culture medium, MTT, dimethyl sulfoxide) were set.

[0061] 3. Experimental results:

[0062] like Figure 3 It was shown that triamterene did not show significant toxicity to HaCaT cells.

[0063] Example 6

[0064] Triamterene combined with Infliximab improves imiquimod-induced psoriasis-like inflammatory lesions in mice:

[0065] Infliximab is an anti-TNF-α monoclonal antibody that has been widely used to treat psoriasis. It can relieve psoriasis symptoms by blocking TNF-α, a key proinflammatory cytokine, and reducing the inflammatory response.

[0066] 1. Experimental animals:

[0067] C57 / B6J female mouse, 68 weeks, 1822 g.

[0068] 2. Experimental methods:

[0069] Take a normal 68-week-old C57 / B6J female mouse and inject 200300μl of anesthetic intraperitoneally according to body weight to anesthetize it. Shave the hair on the back of the mouse to form an exposure area of ​​about 2cm×2cm. Randomly and evenly group them according to body weight. Shaving is performed 12 days before the formal experiment. Weigh the weight of the mouse regularly every day, observe local skin changes, take photos and record them, and give a score of 03 according to the psoriasis area and disease severity (PASI) scoring standard, and the first day is recorded as 0 points. Finally, apply imiquimod cream. Except for the normal control group, other groups apply 5% imiquimod cream 62.5mg / mouse (where the actual content of imiquimod is 3.125mg) on ​​the shaved skin, once a day, for 4 consecutive days. And give corresponding treatment.

[0070] Group 1: normal group, blank control, without any treatment;

[0071] Group 2: Model group, given imiquimod;

[0072] Group 3: Infliximab administration group, given imiquimod and intraperitoneal injection of 10 mg / kg Infliximab.

[0073] Group 4: the triamterene-administered group, which was given imiquimod and 10 mg / kg triamterene by intraperitoneal injection.

[0074] Group 5: Infliximab and triamterene combined administration group, given imiquimod, and intraperitoneal injection of 10 mg / kg triamterene and 10 mg / kg Infliximab.

[0075] The normal group and the model group were given an equal amount of PBS, and the drug group was given 10 mg / kg by intraperitoneal injection according to the dose.

[0076] After 4 days, the skin of the mouse back was taken and photographed to record the condition. The skin lesions were removed and the subcutaneous fat tissue was removed, and the skin was fixed in 4% paraformaldehyde solution for the preparation of paraffin sections.

[0077] 3. Evaluation method:

[0078] The following mouse PASI criteria were established with reference to the PASI (Psoriasis area and severity index) standard to evaluate clinical severity: erythema; infiltration; scaling. Each feature was evaluated with a 0-3 score: 0 = none; 1 = mild; 2 = moderate; 3 = severe. Each site was scored according to the following three clinical characteristics of skin lesions: erythema (E): red or dark red inflammatory plaques that fade when pressed. 0-no erythema is visible; 1-light red; 2-red; 3-very deep red. Infiltration (I): the tendency of the lesion to spread to the surrounding areas, with unclear boundaries and a sense of substance when pressed. 0-the lesion is flush with the normal skin; 1-the lesion is slightly higher than the surface of the normal skin; 2-moderate elevation, the edge of the plaque is round or sloped; 3-the lesion is highly thickened and the elevation is very obvious. Epidermal desquamation / scaling (desquamation, D): refers to the shedding of epidermal cells in pieces. 0 - no visible scales on the surface; 1 - some lesions are covered with scales, mainly fine scales; 2 - most lesions are completely or incompletely covered with scales, and the scales are flaky; 3 - all lesions are covered with scales, and the scales are very thick and layered. Exudation, dryness and itching are not included in the total score.

[0079] 4. Analysis of pharmacological experimental results:

[0080] like Figure 4 The results showed that intraperitoneal injection of 10 mg / kg of triamterene or 10 mg / kg of infliximab alone can significantly improve the psoriasis-like inflammatory lesions induced by imiquimod, and the combined use of triamterene and infliximab can further improve the above lesions.

[0081] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions or improvements made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. Use of a compound of triamterene or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating and / or preventing inflammatory skin diseases.

2. The use according to claim 1, characterized in that The pharmaceutically acceptable salt compound of triamterene is one or more of triamterene nicotinate, triamterene dihydrate, and furosemide-triamterene salt.

3. The use according to claim 2, characterized in that: The inflammatory skin disease is selected from the group consisting of psoriasis, acne, seborrheic dermatitis, pityriasis rosea, cutaneous small vessel vasculitis, perioral dermatitis, tinea capitis, tinea pedis and tinea corporis.

4. The use according to claim 3, characterized in that The inflammatory skin disease is psoriasis.

5. The use according to claim 4, characterized in that The preparation type of the drug is selected from any one of tablets, capsules, granules, micropills, dripping pills, oral liquids, soft capsules, water injections, powder injections, and infusions.

6. Use of a combination comprising one or more compounds selected from the group consisting of triamterene or a pharmaceutically acceptable salt thereof, a TNF antagonist, in the preparation of a medicament for treating and / or preventing inflammatory skin diseases.

7. The use according to claim 6, characterized in that The inflammatory skin disease is psoriasis.

8. The use according to claim 7, characterized in that The TNF antagonist is Infliximab.

9. The use according to claim 8, characterized in that The dosage of triamterene is 1-30 mg / kg, preferably 10 mg / kg; the dosage of infliximab is 10 mg / kg.

10. The use according to claim 9, characterized in that The preparation type of the drug is selected from any one of tablets, capsules, granules, micropills, pellets, oral liquids, soft capsules, water injections, powder injections, infusions, ointments, gels, and microemulsions.

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