Application of multi-target melanocortin receptor stimulant in preparing medicine for preventing and treating pemphigus

Abstract

The invention provides application of a multi-target melanocortin receptor stimulant in preparation of a medicine for preventing and treating pemphigus, and belongs to the technical field of medicines. The invention provides application of a compound WP302 in preparation of drugs for prevention, treatment or adjuvant treatment of pemphigus. The compound WP302 is a good MC1R, MC3R and MC4R multi-target agonist, plays a role through multiple pathophysiological mechanisms, not only has a remarkable curative effect in the aspects of oxidation resistance and inflammation resistance, but also can regulate immune response and promote tissue healing, and a brand new treatment strategy is provided. The medicine is expected to overcome the limitation of the existing treatment method, provides a safer and more effective treatment choice for pemphigus patients, and remarkably reduces long-term side effects.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to the application of a multi-target melanocortin receptor agonist in the preparation of a drug for preventing and treating pemphigus. Background Art

[0002] Pemphigus is a rare autoimmune disease with a high disability rate and potential fatality. It is mainly manifested by large-area blisters and erosions on the skin and mucous membranes, causing great physical and psychological pain to patients and seriously affecting their quality of life. The disease usually affects middle-aged and elderly people, but it may also affect young people. The symptoms of pemphigus in the early stages of the disease are often easily mistaken for common skin diseases, such as eczema or contact dermatitis, but as the disease progresses, the blisters will spread throughout the body, and the patient's skin will experience severe erosion and shedding, which will then cause complications such as infection and malnutrition, and may even be life-threatening.

[0003] Although the exact cause of pemphigus has not yet been fully elucidated, current research has shown that the combined effects of genetic factors, immune system abnormalities, and the external environment play a vital role in the occurrence and development of the disease. In particular, IgG autoantibodies against desmoglein (Dsg1 and Dsg3) play a key role in the pathological mechanism. These autoantibodies attack and destroy the connecting structures (desmosomes) between epidermal keratinocytes, resulting in the loss of the adhesion function of keratinocytes, the loss of the skin's protective barrier function, and ultimately the formation of blisters.

[0004] In the immunopathological mechanism of pemphigus, antibodies activate multiple intracellular signaling pathways by binding to Dsg1 and Dsg3, ultimately leading to epidermal cell separation, i.e., acantholysis. This cell lysis process directly leads to the formation of blisters and causes extensive skin and mucosal damage. As an autoimmune disease, pemphigus is chronic and recurrent, making it difficult to completely cure the disease.

[0005] At present, the progression of pemphigus is mainly controlled clinically through systemic glucocorticoids and immunosuppressants. Although these drugs can effectively relieve symptoms and reduce the formation and spread of skin blisters in the short term, long-term use brings many problems. The side effects of glucocorticoids include weight gain, hypertension, diabetes, osteoporosis, etc., while immunosuppressants may lead to serious adverse reactions such as increased risk of infection and liver and kidney damage. In addition, the long-term control and management of pemphigus faces huge challenges. Nearly half of the patients still need to continue to use glucocorticoids to control the disease after three years of treatment. This long-term reliance on hormones and immunosuppressants for treatment not only increases the patient's living burden, but also increases the economic cost of treatment and the patient's psychological pressure.

[0006] At present, there is no specific drug for the pathological mechanism of pemphigus in clinical practice. The limitations of existing therapies have prompted researchers to actively explore new treatment approaches. In recent years, melanocortins, as an important class of neuropeptide hormones, have gradually attracted widespread attention for their role in regulating inflammatory responses, immune balance and tissue repair. Melanocortins can exert powerful anti-inflammatory, immunomodulatory and healing effects by binding to their specific receptors (such as MC1R and MC5R). In particular, α-melanocortin (α-MSH) has been shown to show good therapeutic potential in a variety of inflammatory and autoimmune skin diseases by inhibiting the expression of multiple proinflammatory factors (such as TNF-α, IL-1, IL-6).

[0007] In addition, melanocortin receptor agonists also play an important role in regulating oxidative stress. Oxidative stress is one of the important pathogenic factors in autoimmune diseases such as pemphigus, and excessive oxidative stress reactions can aggravate the damage of skin cells. Through the activation of melanocortin receptors, this type of agonist can effectively reduce the cell damage caused by oxidative stress, thereby reducing the pathological changes of the skin. In short, the treatment regimen based on the melanocortin system not only has a wide range of anti-inflammatory effects, but also can regulate the imbalance of the immune system and reduce the pathological damage of autoimmune diseases. Therefore, melanocortin receptor agonists provide a new and promising strategy for the treatment of pemphigus, which is expected to fill the gaps in existing therapies, provide more effective treatments, and reduce patients' long-term dependence and the occurrence of side effects. Summary of the invention

[0008] In view of the above problems, the present invention provides the use of a multi-target melanocortin receptor agonist in the preparation of a drug for the prevention and treatment of pemphigus. The present invention provides the use of compound WP302 in the preparation of a drug for the prevention, treatment or adjuvant treatment of pemphigus. The compound WP302 is a good MC1R, MC3R, MC4R multi-target agonist, which works through multiple pathophysiological mechanisms, not only has significant therapeutic effects in anti-oxidation and anti-inflammatory, but also can regulate immune response and promote tissue healing, providing a new treatment strategy. The drug is expected to overcome the limitations of existing treatment methods, provide safer and more effective treatment options for pemphigus patients, and significantly reduce long-term side effects.

[0009] In order to achieve the above-mentioned object of the invention, the technical solution of the present invention is as follows:

[0010] In one aspect, the present invention provides a use of a multi-target melanocortin receptor agonist in the preparation of a drug for the prevention, treatment or auxiliary treatment of pemphigus, wherein the multi-target melanocortin receptor agonist is compound WP302, having the following structure:

[0011]

[0012] Specifically, the active ingredient of the drug includes compound WP302 or a pharmaceutically acceptable salt, solvate, hydrate or isomer thereof.

[0013] Specifically, the drug is used for preventing, treating or assisting in treating pemphigus by binding to melanocortin receptors.

[0014] Preferably, the melanocortin receptors include any one or more of MC1R, MC3R, and MC4R.

[0015] Preferably, the drug achieves the purpose of preventing, treating or assisting in the treatment of pemphigus by one or more of the following mechanisms:

[0016] (1) Enhance antioxidant capacity;

[0017] (2) Inhibit inflammatory response;

[0018] (3) Restoring immune balance;

[0019] (4) Reduce antibody levels in the body.

[0020] Preferably, the dosage form of the drug includes but is not limited to: tablets, pills, powders, suspensions, gels, emulsions, creams, granules, nanoparticles, capsules, suppositories, injections, sprays or injections.

[0021] Specifically, the medicine also includes pharmaceutically acceptable excipients.

[0022] Preferably, the pharmaceutically acceptable excipients include: one or more of solvents, diluents, disintegrants, precipitation inhibitors, surfactants, glidants, adhesives, lubricants, dispersants, suspending agents, isotonic agents, thickeners, emulsifiers, preservatives, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, colorants, flavoring agents, sweeteners, ion exchangers, release agents, coating agents, flavoring agents, and antioxidants.

[0023] Specifically, the drug is administered to mammals.

[0024] Preferably, the drug is administered to humans.

[0025] In a second aspect, the present invention provides a pharmaceutical preparation comprising compound WP302.

[0026] Specifically, the pharmaceutical preparation is used for preventing, treating or assisting in treating pemphigus.

[0027] The positive and beneficial effects of the present invention are:

[0028] The present invention provides the use of compound WP302 in the preparation of a drug for the prevention, treatment or adjuvant treatment of pemphigus. The compound WP302 exerts its effects through multiple pathophysiological mechanisms by enhancing antioxidant capacity, inhibiting inflammatory response, restoring immune balance, and reducing antibody levels in the body, thereby providing a new treatment strategy. The drug is expected to overcome the limitations of existing treatment methods, provide a safer and more effective treatment option for pemphigus patients, and significantly reduce long-term side effects.

[0029] (1) Enhanced antioxidant capacity: Melanocortin family compounds can upregulate the expression of the Nrf2 signaling pathway and significantly improve the antioxidant defense capacity of cells. Activation of Nrf2 can promote the production of a variety of antioxidant enzymes, reduce cell and tissue damage caused by oxidative stress, especially stress damage in skin tissue, and thus improve the pathological state associated with pemphigus.

[0030] (2) Inhibition of inflammatory response: WP302 activates melanocortin receptor 1 (MCR1), effectively inhibits the release of proinflammatory factors (such as TNF-α and IL-17), and significantly reduces local inflammatory response in the skin. By reducing the generation and spread of inflammatory mediators, WP302 can control acute inflammation associated with pemphigus and reduce tissue damage.

[0031] (3) Restoring immune balance: Melanocortin family compounds can restore the body's immune homeostasis by regulating the functions of T cells and other immune cells. Their effects include reducing overactive immune responses, inhibiting autoimmune effects, and reducing the incidence of blister formation and skin erosion. Through the immune regulation mechanism, the drug restores the immune system to normal and reduces the pathological immune response of pemphigus.

[0032] (4) Reducing antibody levels in vivo: WP302 showed the potential to alter disease progression by reducing antibody levels in pemphigus mice.

[0033] (5) Multi-target therapeutic effect: By enhancing antioxidant capacity, inhibiting inflammatory response and restoring immune balance, afamelanotide effectively fights the pathological process of pemphigus at multiple levels. Its multiple mechanisms of action make it a highly effective therapeutic drug that can significantly alleviate disease symptoms and reduce the risk of long-term recurrence, showing good clinical application prospects. DETAILED DESCRIPTION

[0034] The present invention will be further described in detail below in conjunction with specific examples. The following examples are not intended to limit the present invention, but are only intended to illustrate the present invention. The experimental methods used in the following examples are generally conventional, unless otherwise specified, and the materials, reagents, etc. used in the following examples are commercially available, unless otherwise specified.

[0035] 1. Reagents and instruments

[0036] The reagent information used in the present invention is shown in Table 1, and the instrument information is shown in Table 2.

[0037] Table 1 Reagent information

[0038] Name Article No. Batch No. Manufacturer MPLA M130160 1246298-63-4 aladdin DSG3 Protein 00F2M1001 20231227F001 Biointron Normal Saline H37022749 SD2310230 Shandong Hualu Pharmaceutical Hematoxylin BA4041 C230707 Baso Eosin (aqueous solution) BA4098 C230703 Baso Xylene 1330-20-7 20230905 Yonghua Chemical Absolute Ethanol 64-17-5 20230602 Yonghua Chemical

[0039] Table 2 Instrument information

[0040] Name Model Manufacturer Forced Air Drying Oven DHG-9050A Huitai Instrument Automatic Dehydrator TP1020 Leica Microtome RM2235 Leica Embedding Machine EG1150H Leica Freezing Stage EG1150G Leica Slide Spreader HI1210 Leica Wax Melter XH-90D Premiere Slide Scanner MF43-N MShot

[0041] 2. Test products

[0042] The information of the test products used in the present invention is shown in Table 3.

[0043] Table 3 Test product information

[0044]

[0045]

[0046] The structural formula of WP302 is shown in Formula I:

[0047]

[0048] The structural formula of WP203 is shown in Formula II:

[0049]

[0050] 3. Experimental animals

[0051] (1) Animal information: C57BL / 6 (32 animals), female, 8 weeks old.

[0052] (2) Animal source: C57BL / 6 were purchased from Vital River.

[0053] (3) Animal husbandry: Animals were kept in SPF animal rooms, IVC cages, at a temperature of 20-26°C, a humidity of 40-70%, and a 12-hour light-dark cycle. The animals were fed ad libitum, except when fasting was required.

[0054] This experiment uses C57BL / 6 mice, which are standard animals used in non-clinical drug research and have a large amount of background data. They have a clear genetic background, relatively stable responses to drugs, small individual differences, are easy to raise, are small in size, are easy to operate in experiments, and save test products. They are currently internationally recognized as good animals for drug pharmacokinetic and toxicology tests. There is no other known alternative to live animal experiments in this experiment.

[0055] The animal use method of this experiment was approved by the Institutional Animal Ethics Committee (IACUC). The IACUC approval number is SZHK2024059.

[0056] 4. Data processing

[0057] All data were analyzed using GraphPad Prism 10.1.2 software. The analysis methods used in this paper were One-way ANOVA for single factors and Two-way ANOVA for two factors.

[0058] Example 1 Preparation of test sample

[0059] The preparation of the test sample is completed using aseptic procedures. According to the following steps, dilute the stock solution with solvent to the required concentration under the biosafety cabinet / laminar flow hood. The actual preparation process is recorded in the original record. The test sample is prepared once a week and stored at 2-8℃.

[0060] (1) Preparation of WP302 test sample: The required amount of WP302 is 3.6 mg, which is dissolved in 26.88 mL of PBS to a concentration of 0.13 mg / mL.

[0061] (2) Preparation of WP203 test sample: The required amount of WP203 is 82 mg, which is dissolved in 26.88 mL of PBS to a concentration of 3.05 mg / mL.

[0062] Example 2 Evaluation of WP302 efficacy in vivo

[0063] 2.1 Construction and grouping of animal models

[0064] In the present invention, C57BL / 6 mice were randomly divided into 4 groups, 8 mice in each group, namely normal group, model group, WP302 group and WP203 group, and subcutaneous injection was performed according to the following method for 35 consecutive days, and the first day of administration was recorded as Day 0.

[0065] (1) WP302 group: 1.3 mg / kg WP302 test substance was injected subcutaneously twice a day.

[0066] (2) WP203 group: 30 mg / kg WP203 test substance was administered subcutaneously twice a day.

[0067] (3) Normal group: An equal volume of PBS was injected subcutaneously twice a day.

[0068] (4) Model group: An equal volume of PBS was injected subcutaneously twice a day.

[0069] From Day 0, except for the normal group, all mice were subcutaneously injected with 200 μL of a mixture of 50 μg / mL DSG3 protein and MPLA (DSG3 protein and MPLA volume ratio 1:1) to induce pemphigus symptoms in mice, once every two weeks, for a total of two times. Animals were divided into groups and given medication starting from Day 0.

[0070] 2.2 Effect of WP302 on the body weight of pemphigus mice

[0071] During the experiment, C57BL / 6 mice were weighed twice a week, for a total of 11 times. The weight data of mice before and after the experiment are shown in Table 4. The results showed that the weight of mice in each group was normal.

[0072] Table 4 Mouse body weight data (Mean±SEM, n=8)

[0073]

[0074] 2.3 Clinical scoring

[0075] Clinical scoring criteria: 1 point for the presence of erosive lesions in any independent area, with a total score of 11 points; 1 point for the presence of elongated incisors or canines due to lack of solid food intake, with a total score of 1 point; 1 point for the occurrence of hair loss in any independent area, with a total score of 3 points. The total clinical score is 15 points. Clinical scoring was performed twice a week, for a total of 11 times. The clinical score data of each group of animals before and after the experiment are shown in Table 5.

[0076] Table 5 Clinical score data of mice (Mean±SEM, n=8)

[0077]

[0078] The clinical scoring data of mice showed that the score of the normal group was always 0.00±0.00 during the entire experiment, and no pemphigus symptoms appeared; the score of the model group increased significantly to 0.38±0.18 at the end of the experiment, verifying the successful establishment of the pemphigus model; the mean clinical score of the WP203 group was consistent with that of the model group; and the clinical score of the WP302 group was reduced, which could effectively suppress the symptoms of pemphigus to a certain extent.

[0079] 2.4ELISA assay

[0080] On Day 25 and Day 35, non-anticoagulated whole blood was collected twice to separate about 50 μL of serum, and the DSG3 antibody expression level was detected using an ELISA kit. The results of animal DSG3 antibody ELISA assay are shown in Table 6.

[0081] Table 6 DSG3 antibody ELISA data (Mean±SEM, n=8)

[0082]

[0083] Note: ## in the table represents significant difference compared with the model group (P<0.01), *** represents significant difference compared with the normal group (P<0.0001).

[0084] Day 25, the results of DSG3 antibody ELISA in each group of mice showed that compared with the normal group, the serum DSG3 antibody concentration in the model group was significantly increased*** (P<0.0001), indicating that the model was successfully established. There was no significant difference in the serum DSG3 antibody concentration in the WP302 group and the WP203 group compared with the model group.

[0085] Day 35, the results of DSG3 antibody ELISA of mice in each group showed that compared with the normal group, the serum DSG3 antibody concentration of the model group was significantly increased (P<0.0001), indicating that the model was successfully established. Compared with the model group, there was no significant difference in the serum DSG3 antibody concentration of the WP203 group, and the serum DSG3 antibody concentration of the WP302 group was significantly decreased** (P<0.01), indicating that the test product WP302 had a certain efficacy at a dose of 1.3 mg / kg.

[0086] The present invention uses DSG3 to induce C57BL / 6 mice and successfully establishes a mouse pemphigus model. During the experiment, the weight of the mice continued to grow. WP203 (Pumelatide, PT141), a good agonist of melanocortin receptor, did not show the potential for the treatment of pemphigus, indicating that not all melanocortin drugs have the potential for the treatment of pemphigus. Clinical scores and DSG3 antibody ELISA data results show that the test sample WP302 shows a therapeutic effect at a dose of 1.3 mg / kg.

[0087] The above detailed description is a specific description of one of the feasible embodiments of the present invention, and the embodiment is not intended to limit the scope of the present invention. It should be pointed out that any equivalent implementation or change that does not deviate from the present invention should be included in the scope of the technical solution of the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the attached requirements.

Claims

1. Use of a multi-target melanocortin receptor agonist in the preparation of a drug for the prevention, treatment or auxiliary treatment of pemphigus, characterized in that: The multi-target melanocortin receptor agonist is compound WP302, which has the following structure:

2. The use according to claim 1, characterized in that: The active ingredient of the drug includes compound WP302 or a pharmaceutically acceptable salt, solvate, hydrate or isomer thereof.

3. The use according to claim 1, characterized in that: The drug is used for preventing, treating or assisting in treating pemphigus by combining with melanocortin receptors.

4. The use according to claim 3, characterized in that: The melanocortin receptors include any one or more of MC1R, MC3R, and MC4R.

5. The use according to claim 3, characterized in that: The drug is used to prevent, treat or assist in the treatment of pemphigus through one or more of the following mechanisms: (1) Enhance antioxidant capacity; (2) Inhibit inflammatory response; (3) Restoring immune balance; (4) Reduce antibody levels in the body.

6. The use according to claim 1, characterized in that: The dosage forms of the drug include: tablets, pills, powders, suspensions, gels, emulsions, creams, granules, nanoparticles, capsules, suppositories, injections, sprays or injections.

7. The use according to claim 1, characterized in that: The medicine also includes pharmaceutically acceptable excipients.

8. The use according to claim 7, characterized in that: The pharmaceutically acceptable excipients include: one or more of solvents, diluents, disintegrants, precipitation inhibitors, surfactants, glidants, adhesives, lubricants, dispersants, suspending agents, isotonic agents, thickeners, emulsifiers, preservatives, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, colorants, flavoring agents, sweeteners, ion exchangers, release agents, coating agents, flavoring agents, and antioxidants.

9. The use according to claim 1, characterized in that: The drug is administered to mammals.

10. The use according to claim 9, characterized in that: The drug is administered to humans.

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