Use of compound G414-0147 in preparing a drug for promoting skin wound healing

Abstract

The present invention discloses the use of compound G414-0147 in the preparation of a drug for promoting skin wound healing, wherein the molecular formula of compound G414-0147 is C 25 H 30 FN7, with the structural formula #imgabs0#, is a small molecule drug proposed in the present invention for use in preparing a drug that promotes skin wound healing. The drug, in particular, contains compound G414-0147 or a pharmaceutically acceptable salt thereof as an active ingredient. The drug promotes skin wound healing by promoting epidermal keratinocyte migration. The drug is highly safe, stable, can treat large-area skin wounds, and has low synthesis cost. This drug provides a new approach for the clinical treatment of skin wound healing and has important clinical application value.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to use of compound G414-0147 in preparing a medicine for promoting skin wound healing. Background Art

[0002] Trauma is the leading cause of morbidity and disability worldwide, accounting for 12% of all diseases. Skin trauma is the most common of these injuries, with over 10 million new cases each year. When skin defects are excessive, wound infection is severe, or the patient's underlying health conditions are poor, wound healing often becomes difficult, significantly impacting both the individual patient's quality of life and national healthcare expenditures. In the United States alone, non-healing wounds account for approximately $50 billion in annual healthcare costs, of which nearly $12 billion are due to surgical incisions and trauma, and nearly $7.5 billion are due to burns. However, there is currently no ideal treatment method to effectively promote skin wound healing.

[0003] There are 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: It only provides a closed moist environment for the wound, which is conducive to wound healing, but it cannot directly activate the proliferation and migration abilities of wound cells to accelerate the healing process, and the effect is limited.

[0005] 2. Negative pressure sealed drainage: Intermittent or continuous negative pressure suction is used to discharge liquefied necrotic tissue from the wound, thereby clearing 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 accurate.

[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 and are prone to safety issues such as allergies. In addition, their activity is unstable and easily inactivated, which limits their clinical application.

[0007] 4. Surgical treatment: Skin transplantation / flap transplantation is performed by 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 area and leave large scars. It is not suitable for patients with large skin wounds.

[0008] 5. Cell therapy: It is an emerging treatment technology, but it has the risk of causing tumors and is not safe.

[0009] In summary, commonly used methods such as moist dressings and negative pressure closed drainage indirectly promote wound healing by providing a favorable environment for wound healing, but do not directly promote wound growth. Growth factors, cytokines, and stem cell therapies are all biological products with poor safety and stability, and high production costs, which limit their clinical application. Therefore, there is still a lack of convenient and effective methods to promote skin wound healing.

[0010] G414-0147 is a small molecule compound from the Chemdiv compound library (https: / / www.chemdiv.com) with the molecular formula C 25 H 30 FN7, molecular weight is 447.56, structural formula is Figure 1 As shown, it is a lipophilic molecule with low solubility in water. Currently, there is no research and application report on the pharmacological activity of compound G414-0147. Summary of the Invention

[0011] To overcome the problems that currently commonly used methods such as moist dressings and negative pressure sealed drainage cannot directly promote wound growth, and that biological products such as growth factors, cytokines, and stem cell therapies have poor safety and stability, the present invention aims to provide the use of compound G414-0147 in the preparation of drugs that promote skin wound healing. Through high-throughput screening of the Chemdiv compound library, it was found that the small molecule compound G414-0147 can effectively promote the migration of epidermal keratinocytes, thereby promoting skin wound healing. It is used to prepare a new drug that promotes skin wound healing, and proposes a new clinical treatment approach for promoting skin wound healing.

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

[0013] The present invention provides the use of compound G414-0147 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for promoting skin wound healing. The molecular formula of compound G414-0147 is C 25 H 30 FN7, the structural formula is

[0014]

[0015] Preferably, the promoting of skin wound healing refers to completely healing the formed skin wound or reducing the area of ​​the skin wound.

[0016] Preferably, the pharmaceutically acceptable salt of compound G414-0147 is selected from one or more combinations of hydrochloride, hydrobromide, sulfate, acetate, lactate, tartrate, tannate, citrate, trifluoroacetate, malate, maleate, succinate, p-toluenesulfonic acid or benzenesulfonic acid.

[0017] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of compound G414-0147 and / or a pharmaceutically acceptable salt thereof, wherein the molecular formula of compound G414-0147 is C 25 H 30 FN7, the structural formula is

[0018] Preferably, the concentration of compound G414-0147 in the pharmaceutical composition is 1-100 μM.

[0019] More preferably, the concentration of compound G414-0147 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 into an injection, emulsion, tablet, powder, granule, ointment, liposome or oral solution.

[0021] The present invention also provides a small molecule injection preparation comprising a therapeutically effective amount of compound G414-0147 and / or a pharmaceutically acceptable salt thereof, wherein the molecular formula of compound G414-0147 is C 25 H 30 FN7, the structural formula is

[0022] Preferably, compound G414-0147 and / or a pharmaceutically acceptable salt thereof is used as the active ingredient, and its concentration is 1-100 μM, more preferably 5-20 μM.

[0023] The present invention proposes the use of compound G414-0147 in the preparation of a drug for promoting skin wound healing, particularly a small molecule drug containing compound G414-0147 as an active ingredient. Compared with the prior art, the present invention has the following advantages:

[0024] (1) Compound G414-0147 promotes epidermal keratinocyte migration, thereby promoting the healing of skin wounds.

[0025] (2) Compared with stem cell therapy, the small molecule drugs of the present invention are safer and more stable, thus avoiding the disadvantages of poor safety and the possibility of tumor formation; compared with surgical treatment, they can treat large areas of skin wounds, avoiding the defects of small treatment range, damage to the donor area, and large scars; compared with wound covering with moist dressings and negative pressure closed drainage, they avoid the disadvantages of no clear therapeutic target, low efficiency, and poor effectiveness; compared with growth factors / cytokines, they avoid the disadvantages of easy allergy, poor safety of biological products, unstable activity, and easy inactivation; compared with existing drugs, they have significant cost advantages and low synthesis costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is the molecular structure of compound G414-0147.

[0027] Figure 2 The figure shows the migration ability of human epidermal keratinocyte cell line (HaCaT) treated with compound G414-0147 after wounding in the examples.

[0028] Figure 3 This is the wound healing result of the skin wound healing model promoted by compound G414-0147 in the examples.

[0029] Figure 4 The results of the cytotoxicity test (MTT experiment) of compound G414-0147 in the examples are shown.

[0030] Figure 5 These are the results of liver and kidney function tests on mice after subcutaneous injection of compound G414-0147 in the examples. DETAILED DESCRIPTION

[0031] Below in conjunction with embodiment, the present invention is further described:

[0032] Example 1

[0033] 1. Materials

[0034] The physicochemical properties of compound G414-0147: The compound G414-0147 was purchased from Shanghai Siejie Biopharmaceutical Co., Ltd., with the molecular formula of C 25 H 30 FN7, molecular weight is 447.56, structural formula is Figure 1 As shown, after being injected into the tissue at the edge of the wound, due to its good fat solubility, it is not easily absorbed by the hydrophilic tissue and diffuses into the capillaries to enter the systemic circulation, so it can achieve a local effect without acting on other tissues and organs except the skin wound tissue.

[0035] 2. Experimental Methods

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

[0037] Human epidermal keratinocytes (HaCaT) were seeded in six-well plates. When the cells filled the wells, a scratch was created using a micropipette tip. Following the scratch, HaCaT cells were treated with 5 or 20 μM of compound G414-0147 or a control solvent for 24 hours. The reduction in the distance between cells across the scratch was observed and measured using Image J software.

[0038] 2.2 Establishment of a mouse wound healing model

[0039] The wound healing model was based on a previous study (The mouse excisional wound splinting model, including applications for stem cell transplantation. Nature Protocol. 2013; 8(2): 302-9.). Briefly, 12-week-old C57 / BL6 mice were anesthetized, their backs prepared, and a full-thickness excision wound with a diameter of 8 mm was created on the midline of the back. The panniculus carnosus was removed, and a silicone annular splint was sutured and fixed to the skin around the wound using 4-0 silk suture to prevent skin contraction from closing the wound. Mice were photographed immediately after surgery, on days 7, and 14, and wound area was calculated using Image J.

[0040] 2.3 Compound G414-0147 injection

[0041] On the day the small wound healing model was established, compound G414-0147 was injected subcutaneously around the wound. Mice were randomly divided into two groups: a control group (solvent group) and an experimental group (G414-0147 treatment group), with 6 mice in each group. Injections were performed every other day, with 8 injections around each wound. Each injection point was injected with 10 μL of drug or control solvent. A 34-gauge needle (World Precision Instruments, Sarasota, FL) was connected to a 10 μL NanoFil microsyringe (World Precision Instruments) for injection. The injection was performed until the sample was obtained. The concentration of compound G414-0147 in the injection was 50 μM.

[0042] 3. Experimental Results

[0043] 3.1 Compound G414-0147 promotes epidermal keratinocyte migration

[0044] The migration ability of human epidermal keratinocyte cell line (HaCaT) treated with small molecule compound G414-0147 after scratching Figure 2 As shown in the figure, it can be seen that the migration ability of cells treated with compound G414-0147 was significantly higher than that of the control group (P<0.001).

[0045] 3.2 Compound G414-0147 can promote skin wound healing

[0046] After the wound healing model was established, the wound healing conditions of mice in different groups were recorded on days 0, 7, and 14. The results showed that the wound area of ​​mice in the G414-0147 injection group 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 keratinocytes (HaCaT) were seeded in six-well plates. After incubation for 24 hours with various concentrations (1-50 μM) of G414-0147 or a control solvent, 10 μL of MTT solution (5 mg / mL) was added to each well. Incubation continued for 4 hours, and the culture was terminated. The culture medium was discarded, and 100 μL of DMSO was added to each well to allow the crystals to fully dissolve. The absorbance of each well was measured on a microplate reader at a wavelength of 490 nm.

[0050] MTT assay of human epidermal keratinocyte cell line (HaCaT) Figure 4 As shown, treatment with different concentrations of G414-0147 (1-50 μM) had no obvious toxicity to epidermal keratinocytes.

[0051] 2) Mouse liver and kidney function test

[0052] On the 14th day after the wound healing modeling (i.e., 14 days after the injection of G414-0147), blood was collected from the retroorbital venous plexus of the mice, and then the main indicators of liver and kidney function were detected using the ALT detection kit (Abcam, product number ab285263), AST detection kit (Abcam, product number ab263882), creatinine detection kit (ELISA, product number ml037726), and urea nitrogen detection kit (ELISA, product number ml076478).

[0053] The results of mouse liver function test (ALT, AST) and kidney function test (creatinine, urea nitrogen) are as follows Figure 5 As shown, subcutaneous injection of 50 μM compound G194-0712 into the wound had no significant effect on the liver and kidney functions of mice.

[0054] The above is a preferred embodiment of the present invention, but the present invention should not be limited to the contents disclosed in this embodiment. Therefore, any equivalent or modified implementations that do not depart from the spirit disclosed in the present invention fall within the scope of protection of the present invention.

Claims

1. Use of compound G414-0147 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for promoting skin wound healing, wherein the molecular formula of compound G414-0147 is C 25 H 30 FN7, the structural formula is 。 2. The use according to claim 1, characterized in that The promoting of skin wound healing refers to completely healing the formed skin wound or reducing the area of ​​the skin wound.

3. The use according to claim 1, characterized in that The pharmaceutically acceptable salt of compound G414-0147 is selected from one or more combinations of hydrochloride, hydrobromide, sulfate, acetate, lactate, tartrate, tannate, citrate, trifluoroacetate, malate, maleate, succinate, p-toluenesulfonic acid or benzenesulfonic acid.

4. A pharmaceutical composition, characterized in that It comprises a therapeutically effective amount of compound G414-0147 and / or a pharmaceutically acceptable salt thereof, wherein the molecular formula of compound G414-0147 is C 25 H 30 FN7, the structural formula is 。 5. The pharmaceutical composition according to claim 4, characterized in that The concentration of compound G414-0147 in the pharmaceutical composition is 1-100 μM.

6. The pharmaceutical composition according to claim 5, characterized in that The concentration of compound G414-0147 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 into an injection, emulsion, tablet, powder, granule, ointment, liposome or oral solution.

8. A small molecule injection preparation, characterized in that: It comprises a therapeutically effective amount of compound G414-0147 and / or a pharmaceutically acceptable salt thereof, wherein the molecular formula of compound G414-0147 is C 25 H 30 FN7, the structural formula is 。 9. The small molecule injection preparation according to claim 8, characterized in that Compound G414-0147 and / or a pharmaceutically acceptable salt thereof is used as an active ingredient at a concentration of 1-100 μM.

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