New uses of jak inhibitors and medicaments to promote skin stretch regeneration

Abstract

The application discloses a new application of a JAK inhibitor and a medicine for promoting skin stretch regeneration, and application of the JAK inhibitor in preparation of the medicine for promoting skin stretch regeneration; the medicine for promoting skin stretch regeneration comprises the following components: a JAK inhibitor and a carrier, wherein the carrier comprises sodium hyaluronate and propylene glycol; the application provides a new application of the JAK inhibitor, and the JAK inhibitor can promote skin stretch regeneration, in particular, can promote increase of hair follicle density, increase of epidermis thickness, increase of dermis thickness, reduction of fibrosis degree, reduction of immune infiltration degree and reduction of myofibroblast quantity after skin expansion.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to new applications of JAK inhibitors and drugs that promote skin stretch regeneration. Background Technology

[0002] Skin soft tissue expansion, or simply skin expansion, is a technique that involves placing a skin soft tissue expander under normal skin tissue and injecting liquid into the expander to increase its volume. This expands the local soft tissue, allowing for the repair of tissue or organ defects. Compared to traditional methods of skin tissue defect repair, skin soft tissue expansion can activate the regenerative capacity of the patient's own skin tissue, constructing additional skin tissue with normal structure and function without causing rejection. It also allows for the coverage of defects with continuous tissue of similar texture, color, thickness, and sensory capabilities, leaving smaller scars, offering significant technical advantages. After more than forty years of development, skin soft tissue expansion has become a routine method for tissue repair and reconstruction in plastic and reconstructive surgery.

[0003] Although conventional skin and soft tissue expansion surgery has been widely used clinically, it still has some significant drawbacks. First, the treatment process requires at least two surgeries and a long period of fluid injection expansion, consuming considerable time and effort. Second, conventional expansion can only provide a limited area of ​​skin, insufficient for repairing large-area skin defects. Third, there is a certain risk of complications. A meta-analysis by our team showed that the overall complication rate of skin expansion was 17.44%, with infection being the most common complication at 4.58%, followed by tissue necrosis at 3.65%. While over-expansion can provide a larger area of ​​skin tissue, it reduces skin quality and significantly increases the incidence of complications such as ulceration and infection. Therefore, improving techniques to increase the efficiency of skin expansion, expand the expansion area, and prevent complications has always been a key research focus in plastic and reconstructive surgery.

[0004] In the early stages, researchers attempted to improve the efficiency of skin expansion by applying various chemical substances, including papaverine, cytochrome D, and dimethyl sulfoxide, to the skin. However, the clinical application of these chemicals was greatly limited due to their toxic side effects. Existing adjuvant therapies, such as local injection of mesenchymal stem cells or fat grafting, can improve the quality of expanded skin, but their widespread adoption is still significantly limited by issues such as donor availability, cell survival rate, and cost-effectiveness. Therefore, there is an urgent need to find new, safe, simple, and effective adjuvant treatments to overcome the limitations of skin regeneration and further enhance the effect of skin stretching and regeneration.

[0005] In summary, existing skin regeneration treatments have limitations, necessitating the development of a safe and highly effective innovative strategy to promote skin regeneration. This invention is the first to propose the application of JAK inhibitors through local administration in the field of skin regeneration, aiming to overcome traditional technological bottlenecks and provide new solutions for plastic and reconstructive surgery wound repair and abnormal skin regeneration. Summary of the Invention

[0006] This invention overcomes these difficulties and provides a new application of JAK inhibitors. JAK inhibitors can promote skin stretch regeneration. Specifically, they can promote increased hair follicle density, epidermal thickening, dermal thickening, reduced fibrosis, reduced immune infiltration, and reduced number of myofibroblasts after skin expansion surgery.

[0007] To achieve this objective, the present invention provides the following technical solution:

[0008] In a first aspect, the invention provides the use of JAK inhibitors in the preparation of medicaments that promote skin stretch regeneration.

[0009] In existing technologies, JAK inhibitors are a class of targeted drugs that exert their effects by inhibiting the JAK-STAT signaling pathway. Clinically, they are used to treat a variety of diseases, including myelofibrosis, polycythemia vera, and rheumatoid arthritis. In skin diseases, JAK inhibitors are mainly used to treat mild to moderate atopic dermatitis, vitiligo, severe alopecia areata, and certain types of psoriasis. Therefore, their safety has been extensively validated in clinical practice.

[0010] In this invention, the enhancement of skin regeneration through stretching includes, but is not limited to, adjunctive therapy for soft tissue expansion.

[0011] Preferably, the JAK inhibitor includes a selective JAK1 inhibitor and a non-selective JAK inhibitor; more preferably, the selective JAK1 inhibitor includes utpatinib, and the non-selective JAK inhibitor includes ruxolitinib.

[0012] In this invention, upadacitinib is used.

[0013] In this invention, ruxolitinib, generic name: ruxolitinib, trade name: Jakavi, Jakafi, full name: ruxolitinib, ruxolitinib, ruxolitinib, Jakavi, Jakafi, ruxolitinib.

[0014] Preferably, the stretch skin regeneration includes: in-situ skin regeneration after skin expansion surgery.

[0015] Preferably, the promotion of stretched skin regeneration includes: increased hair follicle density, thickened epidermis, thickened dermis, reduced degree of fibrosis, reduced degree of immune infiltration, and reduced number of myofibroblasts after skin expansion surgery.

[0016] Preferably, the administration method of the drug includes: topical application, subcutaneous injection, intradermal injection, and oral administration.

[0017] Preferably, the dosage forms of the drug include: tinctures, aqueous solutions, emulsions, mixtures, syrups, ointments, suppositories, tablets, water pills, honey pills, capsules, soft capsules, drop pills, injections, powders, granules, patches, and targeted preparations.

[0018] Preferably, the drug is administered at a frequency of no less than twice a day.

[0019] A second aspect of the present invention provides a medicament for promoting skin stretch regeneration, comprising the following components: a JAK inhibitor and a carrier, said carrier comprising sodium hyaluronate and propylene glycol.

[0020] Preferably, the volume ratio of the carrier to the JAK inhibitor is (10-20):1.

[0021] Preferably, sodium hyaluronate and propylene glycol are 1% sodium hyaluronate (w / v) and 20% propylene glycol (v / v), respectively.

[0022] Preferably, the JAK inhibitor includes a selective JAK1 inhibitor and a non-selective JAK inhibitor; more preferably, the selective JAK1 inhibitor includes utpatinib, and the non-selective JAK inhibitor includes ruxolitinib.

[0023] Preferably, the stretch skin regeneration includes: in-situ skin regeneration after skin expansion surgery.

[0024] Preferably, the promotion of stretched skin regeneration includes: increased hair follicle density, thickened epidermis, thickened dermis, reduced degree of fibrosis, reduced degree of immune infiltration, and reduced number of myofibroblasts after skin expansion surgery.

[0025] Preferably, the administration method of the drug includes: topical application, subcutaneous injection, intradermal injection, and oral administration.

[0026] Preferably, the dosage forms of the drug include: tinctures, aqueous solutions, emulsions, mixtures, syrups, ointments, suppositories, tablets, water pills, honey pills, capsules, soft capsules, drop pills, injections, powders, granules, patches, and targeted preparations.

[0027] Preferably, the drug is administered at a frequency of no less than twice a day.

[0028] A second aspect of the present invention provides a method for combining skin soft tissue expansion surgery to promote in-situ skin regeneration, comprising the following steps:

[0029] S1. A subcutaneous expander was surgically implanted under the healthy skin of the subject;

[0030] S2. Apply JAK inhibitors to the affected areas of skin expansion for local treatment. The medication should be applied at least once every two days.

[0031] S3. The treatment cycle can be determined according to the specific situation of the subject. Generally, each treatment should last no less than 20 days.

[0032] Compared with the prior art, the beneficial effects and significant progress of the present invention are as follows: The present invention provides the application of JAK inhibitors in drugs that promote the regeneration of stretched skin. Experiments have demonstrated that effective JAK inhibitor treatment can enhance the effect of skin stretch regeneration, solving the problem of the lack of simple and easy-to-implement methods to promote in situ skin regeneration. It can be used in the field of plastic and reconstructive surgery for wound repair. Attached Figure Description

[0033] To more clearly illustrate the technical solution of the present invention, the accompanying drawings used in the embodiments of the present invention will be briefly introduced below.

[0034] Obviously, the accompanying drawings described below are only some of the drawings of the embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort, but these other drawings are also within the scope of the drawings required for the embodiments of the present invention.

[0035] Figure 1 This is the method for constructing the mouse head in situ stretch regeneration model according to Example 1 of the present invention;

[0036] Figure 2 Figure C shows the HE staining (Figure A) and Masson staining (Figure B) results of the mouse head in situ stretch regeneration model at different time points, as well as the corresponding statistics.

[0037] Figure 3 Typical HE and Masson staining images (A) and corresponding statistical graphs (B) of Embodiment 2 of the present invention;

[0038] Figure 4 The images shown are (A) and (B) of the immunofluorescence staining in Example 2 of the present invention.

[0039] Figure 5 Typical HE and Masson staining images (A) and corresponding statistical graphs (B) of Embodiment 3 of the present invention;

[0040] Figure 6 The images shown are (A) and (B) of the immunofluorescence staining in Example 3 of the present invention. Detailed Implementation

[0041] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the present invention is further described below in conjunction with specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight. Unless otherwise specified, the experimental materials and reagents used in the following embodiments are commercially available.

[0042] Unless otherwise specified, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. It should be noted that the terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the exemplary embodiments of this application.

[0043] It should be noted that the terms "first," "second," and "third" (if present), etc., in the specification, claims, and accompanying drawings of the embodiments of this invention, are only used to distinguish different objects and not to describe a specific order. Furthermore, the term "comprising" and any variations thereof are intended to cover non-exclusive inclusion. For example, a process, method, system, product, or device that includes a series of steps or units is not limited to the listed steps or units, but may optionally include steps or units not listed, or may optionally include other steps or units inherent to these processes, methods, products, or devices.

[0044] The technical solution of the present invention will now be described in detail with reference to specific embodiments.

[0045] Example 1: Construction of a mouse head stretch model

[0046] Experimental methods

[0047] The animals used in this example were male C57BL / 6 mice, 7-8 weeks old and weighing 22-25 grams. The surgical procedure was performed under sterile conditions in an SPF-grade animal facility. A brief description of the construction process includes:

[0048] 1.1. Anesthetize the mice, and perform local hair removal and alcohol disinfection on the skin from the back of the neck to the upper back;

[0049] 1.2. Make an incision in the skin of the back of the neck and blunt dissection of the subcutaneous tissue at both ends. Insert the expansion bladder of the customized silicone expander into the subcutaneous tissue of the head. Place the injection chamber under the skin of the back and fix it in place.

[0050] 1.3. Initiate tissue expansion by injecting 0.6 mL of PBS into the expander. Then suture the incision. Control mice (CTRL) underwent the same procedure but without the expansion device placement and saline injection, serving as the baseline for comparison.

[0051] 1.4. Increase the injection of 0.3 ml of PBS every four days to ensure that the dilator undergoes a gradual expansion process;

[0052] 1.5 Mice were sacrificed and dissected at different time points after expansion (day 8, marked as EXPD8; day 20, marked as EXPD20; day 36, marked as EXPD36). Skin samples from the expanded head area were collected for paraffin sectioning, followed by hematoxylin and eosin (HE) staining, as well as Masson trichrome staining.

[0053] Experimental results

[0054] like Figure 1 The diagram illustrates the method for constructing an in situ stretch regeneration model of the mouse head. Figure A shows the mouse after head expander implantation and saline injection, including a top view and a side view. The yellow dashed box marks the injection site, and the red dashed box marks the expander sac site. Figure B shows a timeline of model construction, with the day of surgery designated as Day 0.

[0055] like Figure 2 As shown, HE staining (Figure A) and Masson staining (Figure B) results of the skin in a mouse head in situ stretch regeneration model at different stages, along with corresponding statistics (Figure C). The statistical results show that the stretched skin exhibits a basic phenotype of epidermal thickening, dermal thinning, and subcutaneous fat layer thinning.

[0056] Example 2: Application of Non-Selective JAK Inhibitors

[0057] Experimental methods

[0058] All mice used in this experiment were male C57BL / 6 mice, 7-8 weeks old and weighing 22-25 grams. Surgery and drug administration were performed under sterile conditions in an SPF-grade animal facility. To illustrate the efficacy of JAK inhibitors, mice were randomly assigned to experimental and control groups, receiving different treatment regimens respectively.

[0059] 2.1 Preparation of the ointment: Prepare the topical ointment according to the following instructions:

[0060] 1) Carrier: 1% sodium hyaluronate (w / v), 20% propylene glycol (v / v), balance water;

[0061] 2) Drug stock solution: The active ingredient of the drug, Ruxolitinib, is dissolved in DMSO at a certain therapeutically effective dose. The concentration of the drug stock solution mentioned here is 10 mg / mL, and then sterilized by passing it through a 0.22 μm filter membrane.

[0062] 3) Prepare the experimental group ointment according to the mixing ratio of carrier to drug stock solution = 20:1. The ointment should be freshly prepared and stored at -20 degrees Celsius for long-term storage. It can be stored at 4 degrees Celsius or room temperature for short-term storage.

[0063] 4) A control ointment was prepared concurrently, in which the active pharmaceutical ingredient was replaced with DMSO.

[0064] The ointment in the experimental group was labeled RUXO, and the ointment in the control group was labeled CTRL.

[0065] 2.2 A mouse head stretching model was constructed for 6 mice according to the method in Example 1. After the first mouse surgery, the hair in the skin stretching area was shaved off to facilitate subsequent drug application.

[0066] 2.3 Drug Treatment: After surgery, mice were given topical application of a medicated ointment every two days for 20 days, with an interval of at least 36 hours between applications. The experimental group received Ruxolitinib ointment (RUXO), while the control group received a control ointment (CTRL). Simultaneously, a measured amount of PBS was periodically injected into the injection vessel every four days, as described above, to promote traction regeneration of the scalp skin.

[0067] 4) On day 20 post-surgery (EXPD20), mice in both groups were euthanized and dissected. Skin samples were collected from the head expansion area. Half of the samples were used to prepare paraffin sections, which were then stained with hematoxylin and eosin (HE) and Masson staining. The other half were used to prepare frozen sections, which were then stained with immunofluorescence. K14-positive cells were defined as epidermal cells, K10-positive cells as epidermal cells, F4 / 80-positive cells as macrophages, CD45-positive cells as immune cells, and αSMA-positive cells as myofibroblasts.

[0068] Experimental results

[0069] The results are as follows Figure 3-4 As shown, the Ruxo group consisted of mice receiving ruxotinib ointment for skin and head stretching, while the CTRL group consisted of mice receiving control ointment for skin and head stretching. EXPD20 indicates that stretching skin samples were collected from both groups on day 20 post-surgery. Scale bar = 50 μm.

[0070] Figure 3 Image A shows typical HE and Masson staining images, indicating that the skin structure of the drug-treated mice remained intact. Figure 3B represents the corresponding statistical graph. The statistical results show that, compared with control mice, the drug-treated mice exhibited increased epidermal and dermal thickness of stretched skin, and decreased collagen fiber density, the latter suggesting reduced fibrosis.

[0071] Figure 4 A shows an image stained with immunofluorescence. Figure 4 B represents the corresponding statistical graph. Statistical results show that, compared with control mice, drug-treated mice had thicker K10+ epidermal cells and a higher proportion of K10+K14+ positive cells in their epidermal cells, indicating enhanced epidermal proliferation and differentiation capacity; fewer F4 / 80 positive cells and CD45 positive cells, indicating a lower degree of inflammatory infiltration; fewer myofibroblasts, indicating a lower degree of fibrosis.

[0072] Example 3: Application of Selective JAK1 Inhibitors

[0073] The experimental method in this example is the same as in Example 2, except that the active ingredient in the experimental group is replaced with Upadacitinib.

[0074] The results are as follows Figure 5-6 As shown, the Upat group consisted of mice receiving utpatinib ointment for skin and head stretching, while the CTRL group consisted of mice receiving control ointment for skin and head stretching. EXPD20 indicates that stretching skin samples were collected from both groups on day 20 post-surgery. Scale bar = 50 μm.

[0075] Figure 5 Image A shows typical HE and Masson staining images, indicating that the skin structure of the drug-treated mice remained intact. Figure 5 B represents the corresponding statistical graph. The statistical results show that, compared with control mice, the drug-treated mice had increased epidermal thickness and decreased collagen fiber density in their stretched skin, the latter suggesting a reduction in fibrosis.

[0076] Figure 6 A shows an image stained with immunofluorescence. Figure 6 B represents the corresponding statistical graph. The statistical results show that, compared with control mice, the dermal thickness of drug-treated mice remained unchanged, but the thickness of K10+ epidermal cells was higher, and the proportion of K10+K14+ positive cells in epidermal cells was higher, indicating enhanced epidermal proliferation and differentiation capacity; the number of F4 / 80 positive cells and CD45 positive cells was lower, indicating a lower degree of inflammatory infiltration; the number of myofibroblasts was lower, indicating a lower degree of fibrosis.

[0077] In the description process of the above instruction manual:

[0078] The terms "this embodiment," "an embodiment of the present invention," "as shown," "further," and "further improved technical solutions," etc., indicate that the specific features, structures, materials, or characteristics described in the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms are not necessarily directed at the same embodiment or example, and the specific features, structures, materials, or characteristics described can be combined or combined in any suitable manner in one or more embodiments or examples. Furthermore, without causing contradiction, those skilled in the art can combine or combine the different embodiments or examples described in this specification and the features of the different embodiments or examples.

[0079] Finally, it should be noted that:

[0080] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them;

[0081] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention. Non-essential improvements, adjustments or substitutions made by those skilled in the art based on the content of this specification are all within the scope of protection claimed by the present invention.

Claims

1. Use of a JAK inhibitor for the manufacture of a medicament for promoting skin stretch regeneration, characterized in that, The JAK inhibitor is upadacitinib or ruxolitinib.

2. Use according to claim 1, wherein The stretched skin regeneration comprises skin in-situ regeneration after skin expansion.

3. Use according to claim 2, wherein the compound is ###0002### The promotion of the stretched skin regeneration comprises increased hair follicle density, thickened epidermis, thickened dermis, reduced fibrosis, reduced immune infiltration, and reduced number of myofibroblasts after skin expansion.

4. The use according to claim 1, wherein The administration of the drug comprises topical application, subcutaneous injection, intradermal injection, and oral administration.

5. The use according to claim 1, wherein the compound is ###0002### The dosage form of the drug comprises tincture, water, emulsion, mixture, syrup, ointment, suppository, tablet, water pill, honey pill, capsule, drop pill, needle, powder, granule, and patch.

6. The use according to claim 1, wherein The frequency of the drug administration is not less than 2 times per day.

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