Use of cystatin a in the preparation of a medicament for promoting healing of skin wounds

Abstract

The application discloses use of cystatin A in preparation of a medicine for promoting skin wound healing, wherein the cystatin A is a small molecule secretory protein, the English name of which is Cystatin A, and the molecular weight of which is 12 kDa. The application proposes that the cystatin A is used for preparation of the medicine for promoting skin wound healing, in particular, a small molecule medicine taking the cystatin A as an active ingredient. The cystatin A applied locally on a wound can effectively promote skin wound healing, has high safety, is stable, can treat large-area skin wounds, has low synthesis cost, provides a new way for clinical treatment of skin wound healing, and has important clinical application value.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, and specifically relates to the use of cystatin A in the preparation of drugs that promote the healing of skin wounds. Background Technology

[0002] Trauma is the leading cause of disease incidence and disability worldwide, accounting for 12% of all diseases. Among trauma-related injuries, skin wounds are the most common, with over ten million new cases each year. When skin loss is excessive, wound infection is severe, or the patient's underlying condition is poor, wounds often fail to heal, significantly impacting individual patients' quality of life and national healthcare expenditures. In the United States alone, intractable wounds cost approximately $50 billion annually in healthcare, with surgical incisions and traumatic injuries accounting for nearly $12 billion and burns accounting for nearly $7.5 billion.

[0003] Currently, there are various methods to promote skin wound healing, but all have significant limitations. The main methods include: 1. Covering the wound with moist dressings: This only provides a closed, moist environment to the wound, which is beneficial for healing, but it cannot directly activate the proliferation and migration of wound cells to accelerate the healing process, thus its effect is limited. 2. Negative pressure wound therapy: This method uses intermittent or continuous negative pressure suction to drain liquefied necrotic tissue from the wound, achieving the purpose of removing necrotic tissue and accelerating wound healing. This method indirectly provides a good environment for wound healing, but it does not directly promote wound growth, resulting in limited clinical efficacy and an unclear treatment target. 3. Surgical treatment: Skin grafting / flap transplantation involves excising the wound and taking healthy skin from other parts of the patient to cover it. This method is very painful and can cause damage to the donor site and leave large areas of scarring, and it is not suitable for patients with large skin wounds. 4. Stem cell therapy: This is an emerging treatment technology, but it carries the risk of tumor formation and has poor safety. In summary, commonly used methods such as moist dressings and negative pressure wound therapy indirectly promote wound healing by providing a favorable environment, but they do not directly promote wound growth. Stem cell therapy, on the other hand, suffers from poor safety and stability, and its high preparation cost limits its clinical application. Therefore, convenient and effective methods for promoting skin wound healing are currently lacking.

[0004] Cystatin A, a member of the cystatin superfamily 1, is a protein that functions as a cysteine ​​protease inhibitor, regulating the activity of papain family cysteine ​​proteases such as cathepsin B, H, and L. Current research indicates that cystatin A is one of the precursor proteins of the keratinization capsule of keratinocytes, playing a role in epidermal development and maintenance. Furthermore, cystatin A plays a crucial role in epidermal adhesion in the lower layer of the human epidermis, and loss-of-function mutations in its gene have been shown to be a potential genetic cause of exfoliative ichthyosis. However, the role of cystatin A in promoting skin wound healing has not yet been reported. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the purpose of this invention is to provide the use of cystatin A in the preparation of drugs that promote skin wound healing. It has been discovered that cystatin A plays an important role in the skin wound healing process. Experiments have demonstrated that local application of cystatin A to wounds can effectively promote skin wound healing, overcoming the limitations of commonly used methods such as wet dressings and negative pressure wound therapy in directly promoting wound growth, and the poor safety and stability of biological products such as growth factors, cytokines, and stem cell therapy. This invention proposes a new approach to promote skin wound healing.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] This invention provides the use of cystatin A in the preparation of medicaments that promote the healing of skin wounds.

[0008] Preferably, the dosage form of the drug includes tablets, powders, granules, capsules, oral liquids, injections, or sustained-release formulations.

[0009] Preferably, promoting skin wound healing means that the skin wound formed by the treatment is completely healed or the area of ​​the skin wound is reduced.

[0010] The present invention also provides a small molecule injectable formulation with cystatin A as the active ingredient, the concentration of which is 1-100 μg / mL.

[0011] Preferably, the small molecule injectable formulation uses cystatin A as the sole active ingredient and also includes pharmaceutically acceptable excipients or auxiliary ingredients, wherein the concentration of cystatin A is 1-100 μg / mL.

[0012] This invention proposes the use of cystatin A in the preparation of drugs that promote skin wound healing, particularly small molecule drugs with cystatin A as the active ingredient. Compared with the prior art, the advantages of this invention are:

[0013] (1) Cystatin A can be applied locally to the wound to promote the healing of the skin wound.

[0014] (2) Compared with stem cell therapy, the drug of the present invention has high safety, stable drug, and effectively avoids the disadvantages of poor safety and possible tumor formation.

[0015] (3) Compared with surgical treatment, the drug of the present invention can treat large areas of skin wounds, effectively avoiding the defects such as small treatment range, damage to the donor site and large area of ​​scarring.

[0016] (4) Compared with wound covering with wet dressings and negative pressure closure drainage, the drug of the present invention effectively avoids the disadvantages of no clear treatment target, low efficiency and poor effectiveness.

[0017] (5) It has a significant cost advantage over existing drugs, with lower synthesis costs, providing a new potential approach for clinical treatment of skin wound healing and having important clinical application value. Attached Figure Description

[0018] Figure 1 This is the molecular structure of cystatin A.

[0019] Figure 2 The images show the wound healing status of different groups of mice on days 0, 3, 5, 7, and 10 after the wound healing model was established in this example. Detailed Implementation

[0020] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the described embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0021] Example 1

[0022] 1. Experimental Materials

[0023] Cystatin A was purchased from R&D Systems (Catalog Number: 1407-PI). It is a small secretory protein with the English name Cystatin A (CSTA) and a molecular weight of 12 kDa. Its molecular structure is as follows: Figure 1 As shown (source: Proteindata bank (http: / / www.rcsb.org / pdb / home / home.do), PDB ID:1GD4).

[0024] 2. Experimental Methods

[0025] 2.1 Preparation of Cystatin A Solution

[0026] The cystatin A used in the experiment was dissolved in sterile buffer (containing 25 mM Tris and 100 mM NaCl), and the pH was adjusted to 7.5 to prepare a solution with a concentration of 100 μg / mL.

[0027] 2.2 Establishment of a mouse wound healing model

[0028] The wound healing model was based on previous literature (Inhibiting the cytosolic function of CXXC5 accelerates diabetic wound healing by enhancing angiogenesis and skin repair. Exp Mol Med. 2023 Online ahead of print.). In short, 12-week-old C57BL / 6 mice were anesthetized, and their backs were prepared. An 8 mm diameter full-thickness skin excision was created along the midline of the back, and the dartos fascia was removed. A silicone ring splint was sutured to the skin around the wound using 4-0 silk sutures to prevent skin contraction from causing wound closure. Mice were photographed immediately post-surgery and on days 3, 7, 10, and 14, and the wound area was statistically analyzed using ImageJ.

[0029] 2.3 Cystatin A injection

[0030] On the day the small wound healing model was established, cystatin A injections were started, injected subcutaneously around the wound. Mice were randomly divided into a control group (solvent group) and an experimental group (cystatin A administration group), with 8 mice in each group. Injections were given every 3 days, with 5 injection points around each wound. 20 μL of the drug or control solvent was injected at each point, for a total of 100 μL each time. A 34-gauge needle (World Precision Instruments, Sarasota, FL) connected to a 20 μL NanoFil micro-syringe (World Precision Instruments) was used for injection until the tissue sample was collected. The concentration of cystatin A in the prepared injection solution was 100 μg / mL.

[0031] 3. Experimental Results

[0032] Wound healing in mice of different groups was recorded by photograph on days 0, 3, 5, 7, and 10 after the establishment of the wound healing model. The results showed that the wound area in the cystatin A-treated group was significantly smaller than that in the control group on days 3, 5, 7, and 10 (NS, *P<0.05, ***P<0.001). Figure 2 This study demonstrates that cystatin A can promote skin wound healing.

[0033] The above description is merely a preferred embodiment of the present invention, but the present invention should not be limited to the content disclosed in this embodiment. Therefore, any equivalent or modified versions made without departing from the spirit of the present invention fall within the scope of protection of the present invention.

Claims

1. Use of cystatin A in the preparation of drugs that promote skin wound healing; Promoting skin wound healing refers to the complete healing of skin wounds caused by disease treatment or a reduction in the area of ​​skin wounds.

2. The use according to claim 1, characterized in that, The dosage forms of the drug include powder and injection.

Images