Application of Z-IEED-FMK in preparation of medicine for treating acne vulgaris

Abstract

The invention discloses an application of Z-ITED-FMK in preparation of a medicine for treating acne vulgaris, the Z-ITED-FMK can relieve auricle skin lesion of a mouse acne model, reduce auricle skin thickness, improve inflammatory response and reduce C.acnes titer, the Z-ITED-FMK has a treatment effect on AV, and the Z-ITED-FMK inhibits the inflammatory response of acne local skin and reduces the titer of C.acnes in the skin.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to an application of Z-IETD-FMK in preparing a medicine for treating acne vulgaris. Background Art

[0002] Acne vulgaris (AV) is a common chronic inflammatory skin disease involving the pilosebaceous follicle unit. The global incidence of AV is currently about 9.4%, ranking 8th among the most common diseases in the world. 1 Mostly occurs in adolescents 2 The incidence of this disease is about 39.2%. Skin lesions manifest as widespread acne, papules and pustules on the face, chest and back, and even cystic nodules. Some patients suffer from severe inflammatory reactions involving exposed parts, which lead to obvious pain and recurrence of the disease, often leaving large areas of scars, causing patients to have multiple psychological disorders, bringing heavy psychological burden and mental pressure to patients. 3 Currently, the main treatments for AV are topical or oral antibiotics and isotretinoin. However, antibiotic resistance and ineffective treatment strategies are the main factors leading to poor treatment outcomes for AV. 4 Therefore, newer, safer, and more effective methods are urgently needed to prevent the progression of AV.

[0003] Caspase-8 plays a key role in apoptosis signaling pathways through death receptors 5 In addition, caspase-8 regulates pyroptosis 6 Necroptosis 7 Z-IETD-FMK is a caspase-8 specific inhibitor that inhibits the activation of caspase-8 by inhibiting the cleavage of caspase-8 precursor. 8 The cornea on the surface of the eye is the most innervated tissue in the body and is also the most sensitive tissue, especially sensitive to the stimulation of chemical substances. 9 In previous studies, Z-IETD-FMK was used for local treatment of the ocular surface of the alkali-burned mouse corneal neovascularization model without the side effect of corneal irritation. 10,11 ,These results indicate that Z-IETD-FMK is highly safe and can be used for local skin treatment.

[0004] References

[0005] 1.Wang,Y.,Xiao,S.,Ren,J.&Zhang,Y.Analysis ofthe epidemiologicalburden ofacne vulgaris in Chinabased onthe dataofglobalburden ofdisease2019.Front.Med.9,(2022).

[0006] 2.Li,D.et al.The prevalence ofacne in Mainland China:a systematicreview and meta-analysis.BMJ Open 7,e015354(2017).

[0007] 3.Guo,R.,Xuan,W.,He,X.&Xu,K.Safety and efficacy ofCO2 dot matrixlaser combined with platelet-rich plasma on depressed scar after acnevulgaris and influencing factors of its repair effect:A retrospectiveanalysis.Journal ofCosmetic Dermatology 22,850–861(2023).

[0008] 4.Eichenfield,D.Z.,Sprague,J.&Eichenfield,L.F.Management ofAcneVulgaris:A Review.JAMA 326,2055–2067(2021).

[0009] 5.Marino-Merlo,F.et al.Caspase-8is required for HSV-1-inducedapoptosis and promotes effective viral particle release via autophagyinhibition.Cell Death Differ 30,885–896(2023).

[0010] 6.Wu,J.et al.TNF-αcontributes to sarcopenia through caspase-8 / caspase-3 / GSDME-mediated pyroptosis.Cell Death Discov 9,76(2023).

[0011] 7.Rodriguez,D.A.et al.Caspase-8and FADD prevent spontaneous ZBP1expression and necroptosis.Proc Natl Acad Sci U S A 119,e2207240119(2022).

[0012] 8.Lentini,G.et al.Caspase-8inhibition improves the outcome ofbacterial infections in mice by promoting neutrophil activation.CR Med4,(2023).

[0013] 9.Pollard,R.E.et al.Induction ofIrritation and Inflammation in a 3DInnervated Tissue Model of the Human Cornea.ACS Biomater Sci Eng 6,6886–6895(2020).

[0014] 10.Tian,Y.et al.Pharmacological Inhibition of Caspase-8SuppressesInflammation-Induced Angiogenesis in the Cornea.Biomolecules 10,210(2020).

[0015] 11. Hua, Summary of the invention

[0016] Technical issues solved:

[0017] The present application aims at the deficiencies of the prior art and solves the technical problems that the current treatment methods for acne vulgaris (AV) are mainly topical or oral antibiotics and isotretinoin, while antibiotic resistance and ineffective treatment strategies are the main factors leading to poor treatment effects of AV. The application provides an application of Z-IETD-FMK in the preparation of a drug for treating acne vulgaris, which is a safer and more effective method to prevent the progression of AV.

[0018] Technical solution:

[0019] To achieve the above objectives, this application is implemented through the following technical solutions:

[0020] Disclosed is an application of Z-IETD-FMK in preparing a medicine for treating acne vulgaris.

[0021] Furthermore, the medicine is a solution, suspension, injection, suppository or spray.

[0022] Furthermore, the drug for treating acne vulgaris is administered via intravenous, transdermal, mucosal or smear routes.

[0023] Furthermore, the drug for treating acne vulgaris is administered once a day, once every two days, once every three days, once a week, once a month or once a year during a period of one day to one year.

[0024] Furthermore, the dosage range of the Z-IETD-FMK is 50 to 100 μL / individual body weight (kg).

[0025] Beneficial effects:

[0026] The present application provides an application of Z-IETD-FMK in the preparation of a drug for treating acne vulgaris, which has the following beneficial effects compared with the prior art:

[0027] 1. Z-IETD-FMK alleviates auricular lesions in the mouse acne model, reduces auricular skin thickness, improves inflammatory response, and reduces C.acnes titer;

[0028] 2. Z-IETD-FMK has a therapeutic effect on AV;

[0029] 3. Z-IETD-FMK inhibits the inflammatory response of local acne skin and reduces the titer of C.acnes in the skin. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is a picture of the auricular lesions in the mouse acne model alleviated by Z-IETD-FMK in this application;

[0031] Figure 2 This is a graph showing that Z-IETD-FMK reduces the thickness of the auricle skin in the mouse acne model;

[0032] Figure 3 This is a diagram showing that Z-IETD-FMK improves the inflammatory response of the mouse acne model;

[0033] Figure 4 Graph showing the effect of Z-IETD-FMK on reducing C. acnes titer in the mouse acne model for this application. DETAILED DESCRIPTION

[0034] The present invention will be described below in conjunction with specific embodiments. The described embodiments are only a part of the embodiments of the present invention rather than all of them, and these embodiments are only used to illustrate the present invention and are not used to limit the protection scope of the present invention, which is defined by the claims.

[0035] The experimental methods described in the following examples are conventional methods unless otherwise specified; the reagents and biological materials described are commercially available unless otherwise specified.

[0036] Z-IETD-FMK is a specific inhibitor of caspase-8; acne vulgaris is a chronic inflammatory skin disease involving the pilosebaceous glands.

[0037] Example 1

[0038] Preparation of Propionibacterium acnes suspension:

[0039] Propionibacterium acnes (ATCC6919) was stored under anaerobic conditions at 4°C. When preparing the suspension, one strain of Propionibacterium acnes was taken and activated and proliferated in brain heart infusion medium at 37°C. The colonies were collected, washed and filtered, and the concentration of Propionibacterium acnes was adjusted to 1×10 8 CFU / 20μl).

[0040] Construction of mouse acne vulgaris model:

[0041] Male ICR mice (6-8 weeks old) were injected with 20 μl of Propionibacterium acnes suspension at the same site on the inner side of both ears. 24 hours later, the ears were obviously thickened, the pores at the hair follicle openings were significantly enlarged, and white contents were visible when squeezed.

[0042] Mouse experimental groups:

[0043] Mice were randomly divided into normal, acne, and acne + Z-IETD-FMK groups. 3.9 μg Z-IETD-FMK was dissolved in 100 μl 10% DMSO, with a final concentration of 60 μM. Mice in the acne + Z-IETD-FMK group were injected with Propionibacterium acnes suspension and 100 μl Z-IETD-FMK was applied topically to the skin twice a day for 7 days starting from the day of modeling.

[0044] Observation of changes in mouse auricle skin lesions and measurement of auricle thickness:

[0045] The inflammation, redness, swelling, papules and other skin lesions of the right ear auricle of each group of mice before and after modeling and drug administration were observed with naked eyes and photographed. The thickness of the right ear auricle of mice 7 days after modeling was measured and recorded using a vernier caliper. The same position and strength of the skin lesion were selected for measurement and repeated at least 3 times.

[0046] Hematoxylin-eosin (HE) staining:

[0047] The mouse auricle tissue fixed with 4% paraformaldehyde was routinely dehydrated, paraffin-embedded, sliced, HE-stained, dehydrated with gradient ethanol, transparentized with xylene and sealed with neutral gum, and the pathological changes of the mouse auricle tissue were observed under a microscope.

[0048] Propionibacterium acnes (C.acnes) counts:

[0049] Skin samples (1.5 cm × 2.0 cm) were extracted from the infected site of mice under sterile conditions and placed in a homogenization bottle containing 2.0 ml of PBS solution for homogenization. The homogenate was serially diluted and inoculated onto reinforced clostridial medium (RCM) under anaerobic conditions to count C. acnes, and the log CFU per gram of skin tissue was calculated using the tissue volume.

[0050] Statistical analysis:

[0051] SPSS 22.0 statistical software was used for data analysis; measurement data were expressed as mean ± standard error; one-way ANOVA was used, and P < 0.05 indicated that the difference between the two groups was statistically significant.

[0052] Example 2

[0053] Z-IETD-FMK reduces auricular lesions in mouse acne model:

[0054] The auricles of the mice in the normal group were thin, smooth, and translucent, with clearly visible capillaries; the epidermis of the auricles of the mice in the acne group was red, swollen, rough, and thickened, accompanied by papule-like protrusions; 7 days after Z-IETD-FMK administration, the redness and swelling of the epidermis of the auricles were significantly reduced compared with those in the acne group, most of the papules were absorbed, and the auricles gradually returned to smoothness. Figure 1 shown.

[0055] Figure 1 This is a picture of the auricle skin lesions of the mouse acne model alleviated by Z-IETD-FMK in this application. The mice were randomly divided into normal, acne, and acne + Z-IETD-FMK groups, and the observation results of the ear skin of mice in each group were compared.

[0056] Example 3

[0057] Z-IETD-FMK reduces auricular skin thickness in a mouse acne model:

[0058] Seven days after injection of C. acnes bacterial solution, the auricles of mice in the acne group were significantly thickened compared with those in the normal group (P<0.001). Compared with the acne group, the thickness of the auricle skin in the acne+Z-IETD-FMK group was significantly reduced, and the swelling of the auricle was significantly alleviated (P<0.01).

[0059] Figure 2 This is a diagram showing how Z-IETD-FMK reduces the thickness of the auricle skin in the mouse acne model, and a statistical analysis of the thickness of the mouse auricle skin. *** P<0.001, compared with the normal group, ## P<0.01, compared with the acne group.

[0060] Example 4

[0061] Z-IETD-FMK improves the inflammatory response in the mouse acne model: Figure 3 As shown, Figure 3 This is a diagram of the improvement of the inflammatory response of the mouse acne model by Z-IETD-FMK in this application, and HE staining of the ear skin tissue of each group of mice.

[0062] Z-IETD-FMK reduces C.acnes titers in the mouse acne model:

[0063] The in vivo antibacterial activity of Z-IETD-FMK was evaluated, and the results showed that the number of C. acnes in the acne group was significantly increased compared with the normal group (P<0.001). After topical treatment with Z-IETD-FMK, the number of C. acnes was significantly reduced compared with the acne group, such as Figure 4 shown.

[0064] Figure 4 This is a graph showing the effect of Z-IETD-FMK on reducing the titer of C.acnes in the mouse acne model. This is a statistical analysis of the titer of C.acnes in the ear skin of each group of mice. *** P<0.001, compared with the normal group, ## P<0.01, compared with the acne group.

[0065] The present invention is described above by way of example. It should be noted that, without departing from the core of the present invention, any simple deformation, modification or other equivalent replacement that can be made by those skilled in the art without inventive effort falls within the protection scope of the present invention.

Claims

1. An application of Z-IETD-FMK in the preparation of a drug for treating acne vulgaris.

2. The use of Z-IETD-FMK according to claim 1 in the preparation of a drug for treating acne vulgaris, characterized in that: The medicine is a solution, suspension, injection, suppository or spray.

3. The use of Z-IETD-FMK according to claim 1 in the preparation of a drug for treating acne vulgaris, characterized in that: The drug for treating acne vulgaris is administered via intravenous, transdermal, mucosal or topical routes.

4. The use of Z-IETD-FMK according to claim 1 in the preparation of a drug for treating acne vulgaris, characterized in that: The drug for treating acne vulgaris is administered once a day, once every two days, once every three days, once a week, once a month, or once a year during a period of one day to one year.

5. The use of Z-IETD-FMK according to claim 1 in the preparation of a drug for treating acne vulgaris, characterized in that: The dosage range of the Z-IETD-FMK is 50-100 μL / individual body weight (kg).

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