Use of compound g194-0712 in the manufacture of a medicament for promoting wound healing in skin

Abstract

The application discloses use of a compound G194-0712 in preparation of a medicine for promoting skin wound healing, wherein the compound G194-0712 has a molecular formula of C 20 H 20 N3OCl, and a structural formula. The application provides the compound G194-0712 for preparation of the medicine for promoting the skin wound healing, in particular, a small-molecule medicine taking the compound G194-0712 or a pharmaceutically acceptable salt thereof as an active ingredient, and the effect of promoting the skin wound healing is achieved by promoting epidermal keratinocyte migration; the medicine is high in safety, stable, can treat large-area skin wounds, low in 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 compound G194-0712 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 defects are large, wound infections are 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. However, there is currently no ideal treatment to effectively promote skin wound healing.

[0003] There are currently many methods to promote skin wound healing, but they all have obvious limitations. The main methods are:

[0004] 1. Covering the wound with a moist dressing: This only provides a closed, moist environment for the wound, which is beneficial for wound healing, but it cannot directly activate the proliferation and migration of wound cells to accelerate the healing process, so the effect is limited.

[0005] 2. Negative pressure wound therapy: This method uses intermittent or continuous negative pressure suction to drain liquefied necrotic tissue from the wound, thereby removing necrotic tissue and accelerating wound healing. This method indirectly provides a good environment for wound healing, but does not directly promote wound growth. Its clinical efficacy is limited, and the treatment target is not precise.

[0006] 3. Growth factors / cytokines: They can promote the proliferation of granulation tissue to a certain extent, thereby accelerating wound healing. However, growth factors / cytokines are biological products, which can easily cause safety issues such as allergies. In addition, their activity is unstable and easily inactivated, which limits their clinical application.

[0007] 4. Surgical treatment: Skin grafting / flap transplantation involves excising the wound and taking healthy skin from other parts of the patient to cover the wound. This method is very painful and will cause damage to the donor site and leave large areas of scars. It is not suitable for patients with large skin wounds.

[0008] 5. Cell therapy: It is an emerging treatment technology, but it carries the risk of causing tumors and has a poor safety profile.

[0009] 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. Growth factors, cytokines, and stem cell therapies are all biological products with poor safety and stability, and high manufacturing costs, limiting their clinical application. Therefore, convenient and effective methods for promoting skin wound healing are currently lacking.

[0010] G194-0712 is a small molecule compound from the Chemdiv compound library (https: / / www.chemdiv.com), with the molecular formula C. 20 H 20 N3OCl, with a molecular weight of 353.85, has the following structural formula: Figure 1 As shown, it is a lipophilic molecule with low solubility in water. There are currently no reports on the pharmacological activity and applications of compound G194-0712. Summary of the Invention

[0011] To overcome the limitations of commonly used methods such as wet dressings and negative pressure wound therapy in directly promoting wound healing, and the poor safety and stability of biological products such as growth factors, cytokines, and stem cell therapies, this invention aims to provide the use of compound G194-0712 in the preparation of drugs that promote skin wound healing. Through high-throughput screening of the Chemdiv compound library, it was discovered that the small molecule compound G194-0712 can effectively promote the migration of epidermal keratinocytes, thereby promoting skin wound healing. Its use in the preparation of new drugs that promote skin wound healing proposes a new clinical treatment approach for promoting skin wound healing.

[0012] The above-mentioned objective of the present invention is achieved through the following technical solution:

[0013] This invention provides the use of compound G194-0712 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for promoting skin wound healing, wherein the molecular formula of compound G194-0712 is C 20 H 20 N3OCl, structural formula is

[0014]

[0015] Preferably, promoting skin wound healing means that the existing skin wound is completely healed or the area of ​​the skin wound is reduced.

[0016] Preferably, the pharmaceutically acceptable salt of compound G194-0712 is selected from one or more combinations of hydrochloride, hydrobromide, sulfate, acetate, lactate, tartrate, tannate, citrate, trifluoroacetate, malate, maleate, succinate, p-toluenesulfonic acid, or benzenesulfonate.

[0017] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of compound G194-0712 and / or a pharmaceutically acceptable salt thereof, wherein the molecular formula of compound G194-0712 is C 20 H 20 N3OCl, structural formula is

[0018] Preferably, the concentration of compound G194-0712 in the pharmaceutical composition is 1-100 μM.

[0019] More preferably, the concentration of compound G194-0712 in the pharmaceutical composition is 5-20 μM.

[0020] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient, and can be prepared as an injection, emulsion, tablet, powder, granule, ointment, liposome, or oral liquid.

[0021] The present invention also provides a small molecule injectable formulation comprising a therapeutically effective amount of compound G194-0712 and / or a pharmaceutically acceptable salt thereof, wherein the molecular formula of compound G194-0712 is C 20 H 20 N3OCl, structural formula is

[0022] Preferably, compound G194-0712 and / or its pharmaceutically acceptable salts are used as the active ingredient at a concentration of 1-100 μM, more preferably 5-20 μM.

[0023] This invention proposes the use of compound G194-0712 in the preparation of drugs that promote skin wound healing, particularly small molecule drugs using compound G194-0712 as the active ingredient. Compared with the prior art, the advantages of this invention are:

[0024] (1) The compound G194-0712 promotes the migration of epidermal keratinocytes, thereby promoting the healing of skin wounds.

[0025] (2) The small molecule drug of the present invention has high safety and drug stability compared with stem cell therapy, thereby avoiding the disadvantages of poor safety and possible tumor formation; compared with surgical treatment, it can treat large areas of skin wounds, avoiding the defects of small treatment range, donor site damage and large area of ​​scarring; compared with wound covering with wet dressings and negative pressure wound therapy, it avoids the disadvantages of no clear treatment target, low efficiency and poor effectiveness; compared with growth factors / cytokines, it avoids the disadvantages of easy allergy, poor safety of biological products, unstable activity and easy inactivation; compared with existing drugs, it has a significant cost advantage and low synthesis cost. Attached Figure Description

[0026] Figure 1 The molecular structure of compound G194-0712 is shown.

[0027] Figure 2 The migration ability of human epidermal keratinocytes (HaCaT) treated with compound G194-0712 after scratching in this example is demonstrated.

[0028] Figure 3 The results of the skin wound healing model in the example show that compound G194-0712 promotes skin wound healing.

[0029] Figure 4 The results of the cytotoxicity test (MTT assay) of compound G194-0712 in the examples are shown.

[0030] Figure 5 The results of subcutaneous injection of compound G194-0712 into the wound in the examples show the effects on liver and kidney function in mice. Detailed Implementation

[0031] The present invention will be further described below with reference to the embodiments:

[0032] Example 1

[0033] 1. Materials

[0034] Physicochemical properties of compound G194-0712: Compound G194-0712 was purchased from Shanghai Siejie Biomedical Co., Ltd., and its molecular formula is C2. 20 H 20 N3OCl, with a molecular weight of 353.85, has the following structural formula: Figure 1 As shown, after being injected into the wound edge tissue, due to its good lipid solubility, it is not easily absorbed by hydrophilic tissues and diffuses into capillaries to enter the systemic circulation, thus achieving a local effect without acting on other tissues and organs other than the skin wound tissue.

[0035] 2. Experimental Methods

[0036] 2.1 Cell migration ability test (cell scratch assay)

[0037] Human epidermal keratinocyte cell line (HaCaT) was seeded in six-well plates. When the cells reached confluence, scratches were created using the tip of a micropipette. After scratching, HaCaT cells were treated with 1 μM compound G194-0712, 10 μM compound G194-0712, or a control solvent for 24 hours. The reduction in the distance between cells on both sides of the scratch was observed, and the distance between cells on both sides of the scratch was measured using ImageJ software.

[0038] 2.2 Establishment of a mouse wound healing model

[0039] The wound healing model was based on previous literature (The mouse excisional wound splinting model, including applications for stem cell transplantation. Nature Protocol. 2013; 8(2):302-9.). In short, 12-week-old C57 / BL6 mice were anesthetized, their backs were prepared, and an 8 mm diameter full-thickness skin excision wound was created at the midline of the back, and the dartos fascia was removed. A silicone ring splint was sutured and fixed to the skin around the wound using 4-0 silk sutures to prevent skin contraction from causing the wound to close. The mice were photographed immediately after surgery, on day 7, and on day 14, and the wound area was statistically analyzed using ImageJ.

[0040] 2.3 Injection of compound G194-0712

[0041] On the same day the small wound healing model was established, injections of compound G194-0712 were initiated subcutaneously around the wound. Mice were randomly divided into a control group (solvent group) and an experimental group (G194-0712 administration group), with 6 mice in each group. Injections were administered every other day, at 8 points around each wound. 10 μL of the drug or control solvent was injected at each point using a 34-gauge needle (World Precision Instruments, Sarasota, FL) connected to a 10 μL NanoFil micro-injector (World Precision Instruments). Injections continued until tissue samples were collected. The concentration of compound G194-0712 in the prepared injection solution was 20 μM.

[0042] 3. Experimental Results

[0043] 3.1 Compound G194-0712 can promote the migration of epidermal keratinocytes.

[0044] The migration ability of human epidermal keratinocyte cell line (HaCaT) treated with small molecule compound G194-0712 after scratching was as follows: Figure 2 As shown, the cells treated with compound G194-0712 exhibited significantly higher migration ability than the control group (P < 0.001).

[0045] 3.2 Compound G194-0712 can promote skin wound healing.

[0046] After the wound healing model was established, the wound healing status of mice in different groups was recorded by photograph on days 0, 7, and 14. The results showed that the wound area of ​​mice in the treatment group injected with compound G194-0712 was significantly smaller than that of the control group on days 0, 7, and 14 (P < 0.001). Figure 3 ).

[0047] Example 2

[0048] 1) Drug cytotoxicity test (MTT assay)

[0049] Human epidermal keratinocyte cell line (HaCaT) was seeded in six-well plates. After incubating the cells with different concentrations (1-50 μM) of G194-0712 or control solvent for 24 hours, 10 μL of MTT solution (5 mg / mL) was added to each well, and incubation was continued for 4 hours. The culture was then terminated, the culture medium in the wells was discarded, 100 μL of DMSO was added to each well, and the crystals were allowed to dissolve completely. The absorbance of each well was measured at a wavelength of 490 nm using a microplate reader.

[0050] Results of MTT assay using human epidermal keratinocyte cell line (HaCaT) Figure 4 As shown, treatment with different concentrations of G194-0712 (1-50 μM) had no significant toxicity to epidermal keratinocytes.

[0051] 2) Mouse liver and kidney function test

[0052] On day 14 after the establishment of the wound healing model in mice (i.e., 14 days after the injection of G194-0712), blood was collected from the retro-orbital venous plexus. The main indicators of liver and kidney function were then detected using ALT assay kit (Abcam, catalog number ab285263), AST assay kit (Abcam, catalog number ab263882), creatinine assay kit (ELISA, catalog number ml037726), and blood urea nitrogen assay kit (ELISA, catalog number ml076478).

[0053] Mouse liver function tests (ALT, AST) and kidney function tests (creatinine, blood urea nitrogen) are as follows: Figure 5 As shown, subcutaneous injection of 20 μM MG194-0712 into the wound had no significant effect on liver and kidney function in mice.

[0054] The above description represents 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 compound G194-0712 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for promoting skin wound healing, wherein the molecular formula of compound G194-0712 is C 20 H 20 N3OCl, structural formula is .

2. The use according to claim 1, characterized in that, Promoting skin wound healing refers to the complete healing of existing skin wounds or the reduction of the area of ​​skin wounds.

3. The use according to claim 1, characterized in that, The pharmaceutically acceptable salt of compound G194-0712 is selected from one or more combinations of hydrochloride, hydrobromide, sulfate, acetate, lactate, tartrate, tannate, citrate, trifluoroacetate, malate, maleate, succinate, p-toluenesulfonic acid, or benzenesulfonate.

4. A pharmaceutical composition, characterized in that, It contains a therapeutically effective amount of compound G194-0712 and / or its pharmaceutically acceptable salt, wherein the molecular formula of compound G194-0712 is C 20 H 20 N3OCl, structural formula is .

5. The pharmaceutical composition according to claim 4, characterized in that, The concentration of compound G194-0712 in the pharmaceutical composition is 1-100 μM.

6. The pharmaceutical composition according to claim 5, characterized in that, The concentration of compound G194-0712 in the pharmaceutical composition is 5-20 μM.

7. The pharmaceutical composition according to claim 4, characterized in that, The pharmaceutical composition further comprises a pharmaceutically acceptable carrier and can be prepared as an injection, emulsion, tablet, powder, granule, ointment, liposome, or oral liquid.

8. A small molecule injectable formulation, characterized in that, It contains a therapeutically effective amount of compound G194-0712 and / or its pharmaceutically acceptable salt, wherein the molecular formula of compound G194-0712 is C 20 H 20 N3OCl, structural formula is .

9. The small molecule injectable formulation according to claim 8, characterized in that, The active ingredient is compound G194-0712 and / or its pharmaceutically acceptable salt, at a concentration of 1-100 μM.

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