Use of thiostrepton in the preparation of a medicament for the prevention and treatment of scleroderma

Abstract

This invention relates to the field of pharmaceutical technology, and particularly to the application of thiostrepton in the preparation of drugs for the prevention and treatment of scleroderma. The application of thiostrepton provides a new treatment approach for a wide range of scleroderma patients, offering clinicians and patients more options. For scleroderma patients intolerant to immunosuppressants, thiostrepton can provide topical, oral, or injectable treatments, avoiding the toxic side effects of immunosuppressants. Thiostrepton can be chemically synthesized, making it less expensive than biological agents. Furthermore, as a drug, it has been approved for use in animals, with fewer and milder side effects.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to the use of Thiostrapton in the preparation of medicaments for the prevention and treatment of scleroderma. Background Technology

[0002] Scleroderma is a rare and highly complex autoimmune disease, more common in women, with a peak incidence in the fifth decade of life. The exact cause of scleroderma is unclear, but it may be related to environmental factors that trigger genetic changes in the individual. The main symptoms include: in the early stages, inflammatory changes in the skin, accompanied by vascular lesions, decreased immune system function, and failure of multiple internal organs.

[0003] The pathogenesis of scleroderma is complex, and although it appears to involve the interaction of numerous mechanisms and various molecular signaling pathways, it is not fully understood. Immune responses in scleroderma patients involve innate and adaptive immunity, with the prevalence of Th2 cytokines. Macrophage (M) polarization and mixed M1 / M2 circulating progenitor cells are also present. Alterations in both innate and adaptive immune responses play a crucial role in early pathophysiology, including increased and functionally altered inflammatory cells and products in tissues such as the skin, changes in the number and function of circulating immune cells, and the presence of unique serum autoantibodies detected in most patients.

[0004] Due to the complexity and heterogeneity of scleroderma, treatment is difficult, the disease often inevitably progresses, leading to severe disability, failure of one or more affected organs, and frequently death.

[0005] 1. General treatment

[0006] 1.1 Glucocorticoids: Glucocorticoids are not very effective for this condition. They are usually effective in the early stages of skin lesions (edema stage), joint pain, muscle lesions, serositis, and the inflammatory stage of interstitial lung disease. The dosage is prednisone 30-40 mg / day, used for several weeks, and then gradually reduced to a maintenance dose of 5-10 mg / day.

[0007] 1.2 Immunosuppressants: Commonly used ones include cyclophosphamide, cyclosporine A, azathioprine, and methotrexate. Some reports suggest they may be effective for skin lesions; when used in combination with glucocorticoids, they often improve efficacy and reduce the dosage of glucocorticoids. Methotrexate is effective in improving early-stage skin sclerosis, but ineffective for involvement of other organs.

[0008] 1.3 Other

[0009] In recent years, literature has reported the use of various new treatment methods such as relaxin, imatinib, CD20 monoclonal antibody, and TGF-β antibody, but these have not yet been widely adopted and are only considered for refractory patients.

[0010] 2. Symptomatic and supportive treatment

[0011] 3. Other treatments

[0012] In recent years, peripheral hematopoietic stem cell transplantation sorted from CD34 cells has achieved certain results both domestically and internationally. However, it is expensive and carries a high risk of adverse reactions, and is only recommended for refractory patients.

[0013] While there are many effective drugs for treating inflammatory responses and immune dysregulation, the therapeutic effects and side effects of these drugs remain major factors contributing to the poor prognosis of scleroderma patients. Skin lesions and the resulting organ damage are the causes of the chronic morbidity and high mortality rate of scleroderma. Therefore, the treatment of skin lesions in scleroderma is a major challenge we currently face. Summary of the Invention

[0014] In view of this, the present invention provides the application of Thiostrepton in the preparation of drugs for the prevention and treatment of scleroderma, with the aim of providing an effective, safe and reliable drug for scleroderma patients, especially those with skin fibrosis, to address the problems existing in the current treatment of scleroderma.

[0015] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0016] This invention provides the use of Thiostrapton in the preparation of medicaments for the prevention and / or treatment of scleroderma.

[0017] The present invention also provides the use of Thiostrepton in the preparation of drugs that reduce the expression levels of COL1A1, COL3A1 and / or α-SMA.

[0018] The present invention also provides the use of Thiostrapton in the preparation of medicaments for preventing and / or inhibiting dermal thickening.

[0019] In some specific embodiments of the present invention, the dermal thickening described above includes, but is not limited to, the dermal thickening occurring in scleroderma.

[0020] In some specific embodiments of the present invention, the animal model of scleroderma described above is constructed by bleomycin induction.

[0021] The present invention also provides the use of Thiostrepton in medicaments for the prevention and / or inhibition of skin lesions.

[0022] In some specific embodiments of the present invention, the skin lesions described in the above applications include scleroderma-like lesions.

[0023] In some specific embodiments of the present invention, the animal model of skin lesions described above is constructed by bleomycin induction.

[0024] In some specific embodiments of the present invention, the dosage of the drug in the above applications includes 20 to 40 mg / kg once (equivalent to 2.2 to 4.4 mg / kg for human use), once every two days.

[0025] In some specific embodiments of the present invention, the dosage form of the drug in the above applications includes topical preparations, oral preparations, and / or injectable preparations.

[0026] In some specific embodiments of the present invention, the topical preparations described above include creams, lotions, solutions, gels, ointments, sprays, aerosol sprays, or aerosol foams.

[0027] In some specific embodiments of the present invention, the medicament described in the above applications further includes pharmaceutically acceptable excipients and / or adjuvants.

[0028] In some specific embodiments of the present invention, the above-mentioned excipients include, but are not limited to, binders, fillers, disintegrants, lubricants, isotonicity modifiers, solubilizers, cosolvents, preservatives, colorants, suspending agents, wetting agents, emulsifiers and / or surfactants.

[0029] In some specific embodiments of the present invention, the dosage form of the drug described in the above applications includes a gastrointestinal dosage form or a non-gastrointestinal dosage form;

[0030] The gastrointestinal dosage forms include: powders, tablets, granules, capsules, emulsions, or suspensions;

[0031] The non-gastrointestinal dosage forms include injections, sprays, drops, patches, or ointments.

[0032] In some specific embodiments of the present invention, the dosage form of the above-mentioned drug further includes: liquid dosage form, solid dosage form, semi-solid dosage form or gas dosage form;

[0033] The liquid dosage form includes solutions, lotions, or liniments;

[0034] The solid dosage form includes pills or films;

[0035] The semi-solid dosage forms include gels, suppositories, or pastes;

[0036] The gaseous dosage form includes aerosols.

[0037] The application of this invention has the following effects:

[0038] 1. In the bleomycin-induced scleroderma model, when Thiostrepton was administered at doses of 20 mg / kg and 40 mg / kg, the dermal structure was relatively clear and the fibrotic lesions were milder in the treatment group compared to the model group; the dermal thickness was significantly reduced in the treatment group compared to the model group; when Thiostrepton was administered at a dose of 40 mg / kg, the expression levels of dermal lesion-related factors COL1A1, COL3A1, and α-SMA mRNA were significantly reduced in the treatment group compared to the model group.

[0039] 2. Experimental results show that the minimum effective dose of Thiostrapton in bleomycin-induced scleroderma-like lesions in mice is 20 mg / kg. Attached Figure Description

[0040] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.

[0041] Figure 1 The image shows the results of establishing a bleomycin-induced scleroderma-like skin lesion model in mice in Example 1; where a is the HE and Masson staining map of the successfully constructed skin in the animal model; b is a statistical graph of the skin thickness measurement of the successfully constructed skin in the animal model;

[0042] Figure 2 Example 2 shows the drug prevention experiment of the bleomycin-induced mouse scleroderma skin lesion model treated with Thiostrepton; where a is the skin HE and Masson staining results; b is the skin thickness measurement statistics; c is the mRNA expression level results of lesion-related factors COL1A1, COL3A1, and α-SMA.

[0043] Figure 3 Example 3 shows the drug dosage experiment of Thiostrapton treatment for bleomycin-induced scleroderma skin lesions in mice; where a is a skin HE and Masson staining image; b is a statistical graph of skin thickness measurement. Detailed Implementation

[0044] This invention discloses the application of Thiostrepton in the preparation of drugs for the prevention and treatment of scleroderma. Those skilled in the art can refer to this document and appropriately modify the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can clearly modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0045] This invention provides the use of Thiostrapton in the preparation of medicaments for treating scleroderma.

[0046] Thiostrepton is a cyclic peptide antibiotic composed of thiazole and other structurally interesting groups arranged in a dense array. During protein synthesis, thiorepton exhibits antibiotic activity by inhibiting the dissociation of elongation factor G (EF-G) from the ribosome and the release of inorganic phosphate from EF-G following GTP hydrolysis, thereby blocking ribosome flipping. Thiostrepton has also been described as inhibiting cancer cell growth by downregulating the protein synthesis of the forkhead box M1 (FOXM1) transcription factor (often overexpressed in many malignancies). The binding of the YAP / TEAD / FOXM1 complex to gene regulatory regions controlling the cell cycle may affect cell proliferation.

[0047] Preclinical studies of thiostrepton suggest it can inhibit the growth of human breast cancer and overcome cisplatin resistance. The molecular formula of thiostrepton is C1. 72 H 85 N 19 O 18 S5, with a molecular weight of 1664.89, has the structural formula shown in Formula I.

[0048]

[0049] This patent provides the use of Thiostrapton in the preparation of medicaments for the prevention and treatment of partial or generalized fibrosis of scleroderma.

[0050] Preferably, the drug also contains pharmaceutically acceptable excipients.

[0051] Preferably, the drug is in the form of a topical preparation, an oral preparation, or an injectable preparation.

[0052] As can be seen from the above technical solution, this invention provides the application of Thiostrepton in the preparation of drugs for the prevention and treatment of scleroderma. Scleroderma is a fibrotic lesion affecting part or all of the skin. The technical effects of this invention are as follows:

[0053] Thiostrepton offers a new treatment option for scleroderma patients, providing clinicians and patients with more choices for prevention and treatment. For patients intolerant to immunosuppressants, thiostrepton can provide continued topical, oral, or injectable treatment without the toxic side effects of immunosuppressants. Thiostrepton can be chemically synthesized, making it less expensive than biologics. It has already been approved by the U.S. Food and Drug Administration (FDA) and shows promising clinical application prospects.

[0054] In this invention, the inhibitory effect of Thiostrapton on scleroderma skin lesions was clarified by using a bleomycin-induced scleroderma-like skin fibrosis lesion model.

[0055] In some embodiments, the present invention simulates scleroderma-like skin lesions based on bleomycin-induced skin lesions.

[0056] In some embodiments, the present invention uses literature-recommended concentrations of Thiostreton to treat bleomycin-induced mouse models. Results showed that Thiostreton improved bleomycin-induced cutaneous fibrosis lesions.

[0057] In some embodiments, the present invention uses a mouse model induced by thiostrepton at escalating concentrations. Results showed that the lowest effective dose of thiostrepton to improve bleomycin-induced skin lesions was 20 mg / kg (conversion formula: mouse dose = X mg / kg (human dose) × 70 kg × 0.0026 / 20 g, corresponding to a human dose of 2.2 mg / kg).

[0058] Therefore, Thiostrepton can be used to prevent skin lesions in scleroderma.

[0059] In summary, this invention provides the use of Thiostrapton in the preparation of medicaments for the prevention of scleroderma.

[0060] The drug in question is Thiostrepton.

[0061] Furthermore, the drug also includes pharmaceutically acceptable excipients.

[0062] The drug can be any dosage form currently used in the pharmaceutical industry, including topical preparations, oral preparations, or injectable preparations.

[0063] Each drug dosage form can be prepared by selecting appropriate and acceptable excipients according to the actual needs of that dosage form, which is a conventional dosage form preparation technique in this field. Examples include the preparation of capsules, tablets, and injectable powders.

[0064] Unless otherwise specified, the raw materials, reagents, consumables and instruments used in this invention are all commercially available products and can be purchased from the market.

[0065] The present invention will be further illustrated below with reference to the embodiments:

[0066] Example 1: Establishment of a common animal model of scleroderma (skin)

[0067] I. Materials and Methods

[0068] 1. Bleomycin (BLM)

[0069] Bleomycin was purchased from Xiaoyou Biotechnology Co., Ltd., dissolved in physiological saline, and stored at -80℃.

[0070] 2. Animal model of scleroderma-like skin lesions

[0071] Female C57 mice aged 6–8 weeks (Hunan Slack Jingda Experimental Animal Co., Ltd.) were randomly divided into two groups. The first group was injected subcutaneously with 100 μL of physiological saline in the upper back; the second group was injected subcutaneously with 100 μL of 200 ng / mL bleomycin in the upper back. After 21 days, the mice were treated and their skin tissue samples were collected.

[0072] 3. Experimental detection methods

[0073] HE and Masson staining, RT-PCR (pathogen-associated factor), hydroxyproline detection kit.

[0074] II. Results Analysis

[0075] Figure 1 a. Images of mouse skin sections stained with H&E and Masson staining. The dermal structure of the bleomycin group was more disordered, the fibrosis was more severe, and the thickness was significantly increased compared with the control group, indicating that the bleomycin-induced mouse scleroderma model was successfully established. Figure 1 b (corresponding data are shown in Table 1), a statistical graph of dermal thickness in mouse skin. The bleomycin group showed a significant increase in dermal thickness compared to the control group, indicating that the bleomycin-induced mouse scleroderma model was successfully established. The comparison of dermal thickness between the two groups in the figure was performed using a t-test (*** represents P<0.0010).

[0076] Table 1 Figure 1 b corresponds to the data

[0077]

[0078] Example 2: Thiostrepton can improve bleomycin-induced scleroderma-like lesions in mice.

[0079] I. Materials and Methods

[0080] To investigate the effect of Thiostrepton on bleomycin-induced scleroderma-like lesions, bleomycin-induced scleroderma mice were treated with Thiostrepton at doses of 20 mg / kg or 40 mg / kg (every two days). H&E and Masson staining were used to detect changes in the dermal structure of the mouse skin, and RT-PCR was used to detect changes in the expression levels of factors related to dermal lesions in the mouse skin.

[0081] Female C57 mice aged 6–8 weeks (Hunan Slack Jingda Experimental Animal Co., Ltd.) were randomly divided into 5 groups. Group 1 received a subcutaneous injection of 100 μL of physiological saline in the upper back; Group 2 received a subcutaneous injection of 100 μL of 200 ng / mL bleomycin in the upper back for 8 consecutive days, followed by treatment; Group 3 received a subcutaneous injection of 100 μL of 200 ng / mL bleomycin in the upper back for 8 consecutive days, and simultaneously received a medium dose (20 mg / kg) of Thiostrepton (T); Group 4 received a subcutaneous injection of 100 μL of 200 ng / mL bleomycin in the upper back for 8 consecutive days, followed by a high dose (40 mg / kg) of Thiostrepton. Mice were treated 21 days later, and skin tissue samples were collected.

[0082] II. Results Analysis

[0083] Figure 2 a reflects that in the bleomycin-induced scleroderma model, when Thiostrapton was administered at doses of 20 mg / kg and 40 mg / kg, the dermal structure of the treatment group was relatively clear and the fibrotic lesions were milder compared to the model group. Figure 2 b (corresponding data is shown in Table 2) reflects that in the bleomycin-induced scleroderma model, when Thiostrapton was used at doses of 20 mg / kg and 40 mg / kg, the dermal thickness of the treatment group was significantly reduced compared to the model group. Figure 2 c (corresponding data are shown in Tables 3, 4, and 5) reflects that in the bleomycin-induced scleroderma model, when Thiostrepton was administered at a dose of 40 mg / kg, the expression levels of dermal lesion-related factors COL1A1, COL3A1, and α-SMA mRNA were significantly reduced in the treatment group compared to the model group. The comparison of the three groups in the figure was performed using a two-way ANOVA test (** represents P < 0.01). This suggests that Thiostrepton can improve bleomycin-induced scleroderma-like lesions in mice.

[0084] Table 2 Figure 2 b corresponds to the data

[0085]

[0086] Eight mice were in each group. The solvent control group and the BLM (21 days) group were compared. The BLM (21 days) group was compared with the BLM+T (20 mg / kg) group and the BLM+T (40 mg / kg) group, respectively.

[0087] Table 3 Figure 2 COL1A1 mRNA level data in c

[0088]

[0089] Table 4 Figure 2 COL3A1 mRNA level data in c

[0090]

[0091] Table 5 Figure 2 α-SMA mRNA level data in c

[0092]

[0093] Each group contains 6-7 mice. The solvent control group is compared with the BLM (21 days) group. The BLM (21 days) group is compared with the BLM+T (20 mg / kg) group and the BLM+T (40 mg / kg) group. The values ​​are the mean plus or minus.

[0094] Example 3: The lowest effective dose of Thiostrapton to improve bleomycin-induced scleroderma-like lesions in mice

[0095] I. Materials and Methods

[0096] To determine the minimum effective dose of Thiostrapton to improve bleomycin-induced scleroderma-like lesions in mice, bleomycin-induced scleroderma mice were treated with escalating doses of Thiostrapton (every two days). H&E and Masson staining were used to detect changes in the dermal structure of the mouse skin, and RT-PCR was used to detect changes in the expression levels of factors related to dermal lesions.

[0097] Female C57 mice aged 6–8 weeks (Hunan Slack Jingda Experimental Animal Co., Ltd.) were randomly divided into 5 groups. Group 1 received a subcutaneous injection of 100 μL of physiological saline in the upper back; Group 2 received a subcutaneous injection of 100 μL of 200 ng / mL bleomycin in the upper back; Group 3 received a subcutaneous injection of 100 μL of bleomycin in the upper back and simultaneously administered a low dose (10 mg / kg) of Thiostrepton (T); Group 4 received a subcutaneous injection of 100 μL of bleomycin in the upper back and simultaneously administered a medium dose (20 mg / kg) of Thiostrepton; and Group 5 received a subcutaneous injection of 100 μL of bleomycin in the upper back and simultaneously administered a high dose (40 mg / kg) of Thiostrepton. After 21 days, the mice were treated, and skin tissue samples were collected.

[0098] II. Results Analysis

[0099] Figure 3 a reflects that in the bleomycin-induced scleroderma model, when Thiostrapton was administered at a dose of 20 mg / kg, the dermal structure of the treatment group was relatively clear and the fibrotic lesions were milder compared to the model group. Figure 3 b (corresponding data shown in Table 6) reflects that in the bleomycin-induced scleroderma model, when Thiostrapton was administered at a dose of 20 mg / kg, the dermal thickness in the treatment group was significantly reduced compared to the model group. The comparison of the three groups in the figure was performed using a two-way ANOVA test (** represents P < 0.01). This suggests that the minimum effective dose of Thiostrapton in bleomycin-induced scleroderma-like lesions in mice is 20 mg / kg.

[0100] Table 6 Figure 3 b corresponds to the data

[0101]

[0102] Each group contains 4 or 5 mice. The solvent control group is compared with the BLM group. The BLM group is compared with the BLM+T (10 mg / kg), BLM+T (20 mg / kg) and BLM+T (40 mg / kg) groups, respectively.

[0103] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Use of thiostrepton in the preparation of a medicament for preventing and / or treating scleroderma.

2. Use according to claim 1, wherein The thiostrepton reduces the expression level of COL1A1, COL3A1 and / or α-SMA.

3. The use according to claim 1, wherein The thiostrepton prevents and / or inhibits the thickening of the dermis layer.

4. The use according to claim 3, wherein the compound is ###0002### The thickening of the dermis layer includes, but is not limited to, the thickening of the dermis layer occurring in scleroderma.

5. Use according to claim 1 or 4, wherein the compound is ###0002### The animal model is induced by bleomycin.

6. The use according to claim 1, wherein The dose of the medicament is 2.2-4.4 mg / kg once every two days, calculated as the dose for human.

7. The use according to claim 1, wherein The dosage form of the medicament includes a smearing preparation, an oral preparation and / or an injection preparation.

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