CN116236579A - Application of IL-34 antibody in preparation of medicament for treating psoriasis - Google Patents

Abstract

The invention discloses an application of an IL-34 antibody in preparing a medicament for treating psoriasis. The invention discovers that the psoriasis-like symptoms and pruritus of a psoriasis mouse model can be relieved by inhibiting an IL-34 signal pathway, the characterization and pathological changes of the skin damage area and severity index (PASI) of the psoriasis are obviously improved, the increase of the thickness of the epidermis and the proliferation of lesion skin cells are inhibited, and the inflammatory reaction is inhibited; that is, psoriasis can be treated by inhibiting IL-34 signaling pathway, so that agents inhibiting IL-34 signaling pathway, such as IL-34 antibodies, etc., can be used for the preparation of psoriasis therapeutic drugs. The invention not only widens the application range of the IL-34 antibody, but also enriches the medicines for treating psoriasis, and is beneficial to the treatment of psoriasis patients and the development of novel safe, efficient and economic psoriasis treatment medicines.

Description

Application of IL-34 antibody in preparation of medicine for treating psoriasis

technical field

The invention belongs to the technical field of psoriasis treatment. More specifically, it relates to the application of IL-34 antibody in the preparation of drugs for treating psoriasis.

Background technique

Psoriasis is a common chronic, relapsing, and inflammatory autoimmune skin disease. Its typical skin manifestations are scales and plaques with clear boundaries, and it is easy to recur. Nearly half of the patients are accompanied by discomfort such as itching. According to its clinical manifestations, psoriasis can be divided into four categories: psoriasis vulgaris, arthropathic psoriasis, erythrodermic psoriasis and pustular psoriasis. Among them, psoriasis vulgaris is psoriasis One of the most common types of psoriasis.

At present, the treatment of psoriasis is mainly divided into two types: traditional treatment and targeted treatment of biological agents. Although biological agents or small molecule drugs targeting key cytokines in the treatment mechanism of psoriasis have achieved satisfactory curative effects, they cannot completely cure psoriasis and effectively prevent the recurrence of psoriasis. Poor curative effect and so on. Therefore, it is of great clinical significance to develop safe, efficient and economical new psoriasis therapeutic drugs.

IL-34 (Interleukin 34) is a new cytokine of the interleukin family identified by high-throughput screening of human secreted proteins in 2008 (Lin, H., et al., Discovery of a cytokine and its receptor by functional screening of the extracellular proteome. Science, 2008.320(5877): p.807-11.). After more than ten years of research, although IL-34 has been found to have specific physiological functions and key pathological roles in various diseases, its role in psoriasis has not yet been clarified, and there is currently no target for IL-34. A report on the treatment of psoriasis.

Contents of the invention

The invention provides the application of IL-34 antibody in the preparation of medicines for treating psoriasis, so as to overcome the deficiency of poor curative effect of medicines for treating psoriasis.

The first object of the present invention is to provide the application of the preparation for inhibiting IL-34 signaling pathway in the preparation of psoriasis therapeutic drug.

The second object of the present invention is to provide the application of the preparation inhibiting the IL-34 signaling pathway in the preparation of medicines for alleviating the progression of psoriatic skin lesions.

The third object of the present invention is to provide a medicine for treating psoriasis.

The above object of the present invention is achieved through the following technical solutions:

The present invention analyzes the expression of IL-34 gene and its three receptor genes (Csf1r, Sdc1, Ptprz1) in the lesion skin of patients with psoriasis, adjacent skin and normal human skin, and constructs psoriasis mice Model and detect the expression of IL-34 and its receptor in the skin of psoriasis mouse model, and found that the activation of signaling pathway mediated by IL-34 is involved in the pathogenesis of psoriasis and immune inflammation disorder. At the same time, the present invention inhibits the increase of inflammatory cells and the proliferation of keratinocytes by subcutaneously injecting the IL-34 antibody into the psoriasis mouse model, thereby significantly alleviating the psoriasis Progression of skin lesions in a mouse model. Therefore, the present invention applies for protection of the following applications:

The present invention claims the application of the preparation for inhibiting IL-34 signaling pathway in the preparation of psoriasis therapeutic drug.

Specifically, the psoriasis is psoriasis vulgaris, arthropathic psoriasis, erythrodermic psoriasis or pustular psoriasis.

The present invention also claims the application of the preparation for inhibiting IL-34 signaling pathway in the preparation of medicines for alleviating the progress of psoriatic skin lesions.

Specifically, the relief of the progression of psoriatic lesions is achieved by inhibiting the increase of inflammatory cells, inhibiting the proliferation of keratinocytes and alleviating pruritus symptoms.

Specifically, the psoriasis is psoriasis vulgaris, arthropathic psoriasis, erythrodermic psoriasis or pustular psoriasis.

Specifically, according to the above application, besides the preparation for inhibiting the IL-34 signaling pathway, the medicine also contains its pharmaceutically acceptable adjuvant.

As an optional embodiment, the dosage form of the drug may be an injection or a microneedle.

Specifically, the preparation for inhibiting IL-34 signaling pathway is a preparation for targeting the function of IL-34.

Specifically, the agent for inhibiting IL-34 signaling pathway is IL-34 antibody.

Specifically, the IL-34 antibody is an anti-IL-34 antibody with neutralizing biological activity, that is, an IL-34 neutralizing antibody.

Specifically, the dosage of the IL-34 antibody is at least 5 μg/kg.

The present invention also provides a medicine for treating psoriasis, which contains a preparation for inhibiting IL-34 signaling pathway and pharmaceutically acceptable auxiliary materials thereof.

Specifically, the agent for inhibiting IL-34 signaling pathway is IL-34 antibody.

The present invention has the following beneficial effects:

The present invention found that by inhibiting the IL-34 signaling pathway, psoriasis-like symptoms and itching can be alleviated, and the characterization and pathological changes of psoriasis skin lesion area and severity index (PASI) can be significantly improved, which is specifically reflected in psoriasis Relief and relief of erythema, scales and itching symptoms in the affected area, while inhibiting the increase of epidermal thickness and the proliferation of lesioned skin cells, and inhibiting the inflammatory response, indicating that psoriasis can be improved or treated by inhibiting the IL-34 signaling pathway, which can inhibit IL-34 Preparations of signaling pathways, such as IL-34 antibodies, are used to prepare drugs for treating psoriasis. The invention not only broadens the application range of the IL-34 antibody, but also enriches the drugs for treating psoriasis, and contributes to the treatment of patients with psoriasis and the development of safe, efficient and economical new drugs for treating psoriasis.

Description of drawings

Figure 1 is the analysis of the IL-34 gene and its three receptor genes (Csf1r, Sdc1, Ptprz1) using the GEO database (GSE13355) in clinical psoriasis patients with lesion skin (PP), adjacent skin (PN) and normal people The expression in the skin (NN); NN is 64 cases of normal human skin samples, PN is the skin samples of 58 cases of psoriasis patients, PP is the skin samples of 58 cases of psoriasis patients; the data comparison between multiple groups adopts One -Way ANOVA, Tukey's post hoc test, **P<0.01, ***P<0.001 is the comparison between PP and NN group; ^#### P<0.0001 is the comparison between PP and PN group.

Figure 2 shows the expression of IL-34 and its receptors in the skin of imiquimod-induced psoriasis-like mice (WT IMQ) and Vaseline control mice (WT CON); A in the figure is IL-34 The results of the bulk RNA-seq data of the back skin of the mRNA and its receptor mRNA on the 2.5th day after the establishment of the model in different groups of mice; The expression of IL-34 protein in the back skin on the 2.5th and 4.5th days; C and D in the figure are the CSF1R immunohistochemical staining results and statistical charts of mouse skin tissues in different groups; E in the figure is the model group Immunofluorescence double-staining results of CSF1R in skin lesions on day 4.5 post-implantation; compared using the Mann-Whitney test for two independent samples, *P<0.05, **P<0.01.

Figure 3 shows the treatment results of psoriasis model mice by subcutaneous injection of IL-34 antibody; A in the figure is the back skin image of psoriasis mice on the 4th day after the model is established; B in the figure is the mice of different groups Psoriasis Area and Severity Index (PASI) score statistical graph (left), skinfold thickness change (middle) and time-course graph of touch-induced itching behavior test (Alloknesis, right); Vehi in the figure is the solvent control group, that is, the same amount of petroleum jelly was applied to the back skin and the mouse group was subcutaneously injected with normal saline, and Anti-IL-34 was the control group, that is, the same amount of petroleum jelly was applied to the back skin and the Anti-IL-34 antibody was injected subcutaneously. Vehi+IMQ is the model mouse group in which imiquimod (IMQ) is applied to the back skin after subcutaneously injecting normal saline 30 minutes in advance; After -34, the treated mice group was given IMQ on the back skin; using Two-Way ANOVA analysis, Tukey's post hoc test, ****P<0.0001 is the comparison between Vehi+IMQ and Vehi group; ^### P<0.001, ^#### P<0.0001 is the comparison between Anti-IL-34+IMQ and Vehi+IMQ groups.

Figure 4 shows the effect of IL-34 antibody treatment on the increase of skin inflammatory cells, proliferation of epidermal keratinocytes, and excitability of sensory neurons in psoriasis mice; A in the figure is Vehi+IMQ group and Anti-IL-34+ Immunofluorescent staining samples of macrophages (marker F4/80), CSF1R, cell proliferation (marker Ki67), and neutrophils (marker NIMP-R14) in the skin tissue of mice in the IMQ group; B in the figure It is the statistical chart of the results shown in A, using One-Way ANOVA analysis, Tukey's post hoc test, **P<0.01, ****P<0.0001 is the comparison between Vehi+IMQ group and Vehi group; ^## P<0.01, ^### P<0.001, ^#### P<0.0001 is the comparison between Anti-IL-34+IMQ group and Vehi+IMQ group; C in the figure is shallow dorsal root ganglion (DRG) and spinal dorsal horn (SDH) Changes in neuron excitability (marker p-ERK) in the layer; D in the figure is the statistical graph of the results shown in C; using independent sample t test, ^#### P<0.0001 is Anti-IL-34+ IMQ group compared with Vehi+IMQ group.

Detailed ways

The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the technical field.

Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

Example 1 Expression Changes of IL-34 Gene and Its Receptor

The invention detects the expression changes of the IL-34 gene and its receptor in skin lesions of patients with clinical psoriasis, adjacent skin and normal skin. Specifically, the present invention uses the GEO database to find a biopsy skin total RNA data (GSE13355) covering 58 cases of psoriasis (including lesion skin and adjacent skin) and 64 cases of normal healthy controls. Through platform annotation (GPL570), the expression of IL-34 gene and its three receptor genes (Csf1r, Sdc1, Ptprz1) in normal healthy control samples (NN), paralesional skin samples (PN) and lesioned skin samples of psoriasis patients was analyzed. The expression profile of the sample (PP).

The expression of IL-34 gene and its three receptor genes (Csf1r, Sdc1, Ptprz1) analyzed by GEO database (GSE13355) in clinical psoriasis patients' lesion skin, adjacent skin and normal skin is shown in Figure 1 NN is the skin samples of 64 normal people, PN is the paralesional skin samples of 58 psoriasis patients, and PP is the lesion skin samples of 58 psoriasis patients. It can be seen from Figure 1 that although the expression of IL-34 mRNA in the lesion skin group (PP) of psoriasis patients was not statistically different from that of the normal healthy control group (NN), the parapathic tissue group (PN) of psoriasis patients had no significant difference. The expression of IL-34 mRNA was significantly higher in NN group and PP group. In addition, the CSF1R (Csf1r) mRNA in the receptor of IL-34 was significantly decreased in the lesions and adjacent skin of psoriasis patients, but the mRNA levels of SDC1 (Sdc1) and PTP-ζ (Ptprz1) were significantly lower in the lesion skin group of psoriasis patients. The expression was significantly higher than that in the healthy skin group; the above results suggested that IL-34 may be involved in the occurrence and development of psoriasis.

Example 2 Expression of IL-34 and its receptors in mouse psoriasis-like skin induced by imiquimod

The invention uses imiquimod (IMQ) to induce a mouse psoriasis-like model, and detects the expression of IL-34 and its receptor in the induced mouse psoriasis-like skin.

1. Construction of mouse psoriasis-like model and acquisition of skin tissue samples

The experimental mice used in this example are female C57BL/6J mice, which were purchased from the Animal Experiment Center of Sun Yat-sen University, and were bred in an SPF environment until they were 8-10 weeks old before the animal experiments began. The animal experiments in the present invention have been approved by the Experimental Animal Ethics Committee of Sun Yat-sen University.

The mice were numbered and randomly divided into an IMQ-induced psoriasis-like model group and a Vaseline control group (n=16-18 mice/group). Three days before the start of the experiment, skin was prepared on the back of the mouse with an electric shaver to form an exposed area with a size of 2 × 3 cm ² . Apply 5% imiquimod cream (purchased from Inova Pharmaceuticals, dosage and usage: 10 mg/cm ² , 1 time/day) on the back skin of mice in the psoriasis-like model group for 5 times. On the first day of modeling, light erythema and scales appeared on the mouse skin, and it became the most serious on the 4.5th day, indicating that the psoriasis-like mouse model was successfully established; the mice in the Vaseline control group were externally applied to the back skin, etc. amount of petroleum jelly (con group). On the 2.5th day of modeling (IMQ 2.5d), the back skin tissues of the two groups of mice were collected for subsequent Bulk RNA-seq sequencing analysis. On the 4.5th day of modeling (IMQ 4.5d), the back skin tissues of the two groups of mice were collected for ELISA, immune Histochemistry and immunofluorescence staining.

2. Bulk RNA-seq sequencing and analysis

The back skin tissues of psoriasis-like model mice (IMQ group) and Vaseline control group mice (CON group) after modeling for 2.5 days were sent to Biomarker Biotechnology Co., Ltd. for eukaryotic reference genome RNA For transcriptome analysis (n=4 mice/group), the reference genome was GRCm38_release95, the differential expression analysis software was edgeR_DESeq2, and the screening conditions were FDR<0.05; Fold Change (FC)≥1.5.

The results of bulk RNA-seq data of IL-34 and its receptor mRNA in the back skin of IMQ psoriasis-like model mice (WT IMQ) and Vaseline control mice (WTCON) on day 2.5 are shown in Figure 2 As shown in A, it can be seen from A in Figure 2 that in the skin of psoriasis-like mice, the mRNA expression of IL-34 and its receptor CSF1R decreased significantly, but the other two receptors of IL-34, PTP-ζ (Ptprz1) and Syndecan-1 (Sdc1) mRNA expression levels increased significantly; n=4/group.

3. Detection of IL-34 protein expression level in psoriasis-like model mouse skin lesions and healthy control skin by ELISA

The back skin of the mouse was fully lysed with RIPA lysate (containing 1% protease inhibitor), centrifuged at 10,000 g for 10 minutes at 4° C., and the supernatant was taken. According to the instructions of the ELISA kit (Elabscience, Catalog#: E-EL-M0720C-96T), protein standards were prepared and samples were diluted; 2 to 3 duplicate holes were set up for each standard and sample, and a multichannel pipette was used in the ELISA Standards and samples are added to the 96-well plate of the kit. Incubate the standard and samples according to the conditions recommended in the kit instructions, and then add the primary antibody, HRP, substrate working solution and stop solution in sequence to complete the ELISA operation steps. The absorbance of each well of the 96-well plate was detected by an automatic enzyme-linked immunosorbent analyzer (Bio-Rad Company); the concentration of IL-34 in each skin sample was calculated according to the concentration of the standard substance by fitting a linear working curve.

The results of the content of IL-34 in the skin lesion skin tissue of the psoriasis model mouse obtained by homogenizing the skin sample and performing ELISA detection and the petroleum jelly control skin tissue are shown in B in Figure 2, as can be seen from B in Figure 2, Compared with the Vaseline control group, there was no significant difference in the IL-34 content in the lesions of the psoriasis model mice, n=4-6 mice/group.

4. Detection of protein expression level of CSF1R in psoriasis model mouse skin lesions and Vaseline control skin by immunohistochemistry

The back skin of the mice was collected and fixed in 10% formaldehyde, embedded in paraffin, and sectioned; after dewaxing, hydration, antigen retrieval and removal of endogenous peroxidase, the primary antibody (Anti-CSF1R antibody, Santa Cruz Company, Catalog#: SC-692) and secondary antibody (enhanced enzyme-labeled goat anti-mouse/rabbit IgG polymer, Zhongshan Jinqiao Company, Catalog#: PV-9000), and DAB kit (Vector Laboratories Company, Catalog #:sk-4100) for color development; use a pathological slice scanning imager to scan slides, and measure the gray value of CSF1R in the skin samples of each group by ImageJ software.

The results of CSF1R immunohistochemical staining of mouse skin tissues in different groups and their statistical charts are shown in C and D in Figure 2, respectively, showing the relative fluorescent gray value (RelOD) and cell changes of CSF1R positive signals between groups, n =6/group.

5. Detection of CSF1R expression cell types by immunofluorescence double staining

After 4.5 days of the model, the back skin tissues of each group were collected and fixed in 4% paraformaldehyde. After 4 hours, they were dehydrated in 30% sucrose solution for 2 days until they dehydrated and sank to the bottom in the sucrose solution; the dehydrated specimens were embedded in OTC The tissue was embedded in liquid solution for 5 min, and the tissue was cut into 16-18 μm thin slices with a cryostat, and adhered to an adhesive slide, and left to dry at room temperature overnight; select the dried slides and put them into a washing tank, and shake them slowly with PBS Wash three times, each time for 5 minutes; then use 5% donkey serum blocking solution to slowly shake at room temperature for 1 hour; absorb the blocking solution, add the primary antibody, and put it in a 4°C shaker overnight for 18 hours. Primary antibodies include Anti-CSF1R antibody (Santa Cruz Company, Catalog #: SC-692), Anti-F4/80 antibody (Biolegend Company, Catalog #: 123102), Anti-Iba1 antibody (Abcam Company, Catalog #: ab5076), Anti-CD3 antibody (Biolegend Company, Catalog#: 100214), NIMP-R14 antibody (Santa Cruz Company, Catalog#: SC-59338), Anti-PECAM1 antibody (R&D Company, Catalog#: AF3628); Wash three times with PBS, 15 minutes each time. Then incubate with the corresponding fluorescent secondary antibody (diluted in TBST), and incubate for 1 hour at room temperature with slow shaking in the dark. After absorbing the secondary antibody, wash it three times with PBS, each time for 15 minutes; finally drop the anti-fluorescence quenching mounting medium (with DAPI), slowly cover the coverslip to prevent the generation of air bubbles, and drip a small amount of nail polish around Hold the coverslip. Fluorescent pictures were taken with an ordinary fluorescence microscope (EVOS FL Imaging System), and filming parameters such as exposure time and gain were fixed in the same batch of experiments to ensure that each sample was comparable and reduce experimental errors.

Result analysis: The fluorescence intensity and cell count were analyzed using ImageJ software. The relative integrated density (IntDen) was used to quantify the fluorescence intensity, and ImageJ software was used to circle the position to be measured and the average gray value was automatically calculated.

The immunofluorescent double-staining results of CSF1R in the skin lesions of the model group on the 4.5th day after modeling are shown in Figure 2 E. From Figure 2 E, it can be seen that CSF1R is mainly expressed in macrophages (F4/80, Iba1 mark), but not in T cells (CD3 mark), endothelial cells (PECAM1 mark) and neutrophils (NIMP-R14 mark).

From the above results, it can be seen that although the expression level of IL-34 protein in the skin of vaseline control and psoriasis-like model mice has no significant change, the expression of its receptors has significant changes: CSF1-R, PTP-ζ and Syndecan-1 is abnormally highly expressed in psoriatic lesional skin. Combined with the results in Figure 1, IL-34 is highly expressed in clinical psoriasis adjacent tissues, suggesting that the activation of IL-34-mediated signaling pathways may be involved in the pathogenesis of psoriasis, affecting its phenotype and immune-inflammatory disorders.

The effect of the neutralizing antibody of embodiment 3IL-34 on psoriasis

In order to clarify whether the activation of the IL-34 signaling pathway in the psoriatic lesions is involved in the course of psoriasis, the present invention uses subcutaneous injection of IL-34 in the psoriasis-like mouse model induced by imiquimod (IMQ). Neutralizing antibodies to observe whether inhibiting the IL-34 signaling pathway can effectively treat psoriasis.

1. Grouping of mice

24 female C57BL/6J mice were purchased from the Animal Experiment Center of Sun Yat-Sen University and raised in an SPF environment until they were 8-10 weeks old before the animal experiment. The mice were numbered and randomly divided into 4 groups: vehicle control group (Vehi), simple anti-IL-34 antibody group (Anti-IL-34), psoriasis group (Vehi+IMQ) and anti-IL-34 antibody plus Psoriasis treatment group (Anti-IL-34+IMQ), n=6/group. Three days before the start of the experiment, an electric shaver was used to prepare the skin on the back of the mouse to form an exposed area with a size of 2 × 3 cm ² . The animal experiments in the present invention have been approved by the Experimental Animal Ethics Committee of Sun Yat-sen University.

2. Construction of mouse psoriasis-like model and therapeutic intervention

The modeling process of psoriatic mice is the same as that in Example 2.

An anti-IL-34 antibody with neutralizing biological activity (R&D Company, Catalog#: AF5195) was used to treat psoriasis and anti-IL-34 antibody control group mice. The specific operation was to subcutaneously inject 100 μL of anti-IL-34 antibody at four points on the back 30 minutes before each application of imiquimod cream or petroleum jelly (5 μg/kg, 1 time/day, 5 times in total). The mice in the solvent control group and the psoriasis group were subcutaneously injected with 100 μL of normal saline on the back 30 minutes before cream application every day (1 time/day, 5 times in total). On the 4.5th day of modeling, the gross phenotype of the back skin lesions of the two groups of mice were collected by camera (Fig. 3A), the psoriasis area was measured and the severity index (PASI score) was calculated (Fig. 3B left), and then the skin was measured with a vernier caliper. Changes in pleat thickness (Fig. 3B middle) were then detected by the behavioral test of touch-evoked itching (Alloknesis, Fig. 3B right).

The treatment results of subcutaneous injection of IL-34 antibody on psoriasis model mice are shown in Figure 3; A in Figure 3 is the back skin image of psoriasis mice on the 4th day after modeling; B in Figure 3 is Statistical chart of psoriasis area and severity index (PASI) score (left), skinfold thickness change (middle) and time-course chart of psoriasis behavioral test (Alloknesis, right) in different groups of mice; Vehi is the solvent control group, that is, the mouse group with the same amount of vaseline applied to the back skin and subcutaneously injected with normal saline; IL-34 antibody mouse group; Vehi+IMQ is the model group in which saline was injected subcutaneously 30 minutes in advance and imiquimod was applied to the back skin; Anti-IL-34+IMQ was injected Anti-IL- After 34 hours, the treatment group who applied IMQ to the local skin was given.

It can be seen from Figure 3 that, compared with the vehicle control group (Vehi), the psoriasis model group (Vehi+IMQ, n=6/group) had psoriasis-like dermatitis symptoms, including edema, erythema and scales (Figure 3A) , skin lesion PASI score (Fig. 3B left), pre-model changes in skinfold thickness (Fig. 3B middle), and touch-induced itching behavior (Alloknesis, Fig. 3B right) were all significantly increased. After subcutaneous injection of anti-IL-34 neutralizing antibody, the psoriasis-like skin symptoms of the psoriasis group (Anti-IL-34+IMQ) were significantly improved: such as the reduction of erythema and scales observed by the naked eye (Figure 3A right), PASI score Significantly decreased (Fig. 3B left), skinfold thickness also decreased (Fig. 3B middle); moreover, touch-evoked itch behavior was also significantly improved (Alloknesis, Fig. 3B right). The above results indicate that subcutaneous injection of IL-34 antibody can significantly and effectively alleviate the progression of lesional skin in psoriatic mice.

Example 4

In order to further observe the effects of anti-IL-34 antibody treatment on skin inflammation, skin thickness and pruritus behavior of psoriatic mice, the present invention detected the effect of anti-IL-34 antibody treatment on the increase of inflammatory cells, epidermal keratinocytes, Effects on proliferation and excitability of sensory neurons.

After 4.5 days of modeling, the back skin tissue, dorsal root ganglion (DRG) and spinal dorsal horn (SDH) samples of each group were collected and fixed in 4% paraformaldehyde. The subsequent immunofluorescence experiments were performed as in Example 2. Among them, the newly added primary antibodies include: Anti-Ki67 antibody (Abcam Company, Catalog #: ab16667), p-ERK antibody (Santa Cruz Company, Catalog #: SC-59338).

IL-34 antibody treatment has an effect on the increase of dermal macrophage cells (F4/80 mark), neutrophils (NIMP-R14 mark) inflammatory cells, epidermal keratinocyte proliferation (Ki67 mark) and dorsal root Figure 4 shows the effects of sensory neurons in the ganglion (DRG) and superficial projection neurons in the spinal dorsal horn (SDH). A in Fig. 4 is the macrophage (marker F4/80), CSF1R, cell proliferation (marker Ki67), neutrophil ( Marker NIMP-R14) immunofluorescence staining image; B in Figure 4 is the statistical graph of the results shown in A; C in Figure 4 is the excitability of neurons in the superficial layer of DRG and SDH (marker p-ERK) The change situation; D in Fig. 4 is the statistical chart of the result shown in C.

It can be seen from Figure 4 that macrophages (F4/80, CSF1R markers), cell proliferation (Ki67 markers), neutrophils (NIMP-R14 markers) and neutrophils (marker ) ratio was significantly reduced to the recovery control level, and neuronal excitability (marker p-ERK) in the superficial layer of DRG and SDH was also significantly reduced. Cell molecular level proved that 5 μg/kg anti-IL-34 antibody can effectively alleviate the progression of psoriatic mice skin lesions by inhibiting the increase of inflammatory cells, inhibiting the proliferation of epidermal keratinocytes, and alleviating pruritus symptoms.

It should also be noted that the present invention adopts local injection of skin lesions for administration, which reduces the systemic effect of drug system administration. And as shown in Figure 4, the anti-IL-34 antibody + IMQ group and the anti-IL-34 antibody control group are similar to the solvent control group mice, skin thickness, inflammatory cells and proliferating cells, which indirectly indicates that the administration of anti-IL-34 antibody security. The above results further confirm that subcutaneous injection of anti-IL-34 antibody has the potential to treat psoriasis, and can be used in the preparation of psoriasis therapeutic drugs.

The above-mentioned embodiment is a preferred embodiment of the present invention, but the embodiment of the present invention is not limited by the above-mentioned embodiment, and any other changes, modifications, substitutions, combinations, Simplifications should be equivalent replacement methods, and all are included in the protection scope of the present invention.

Claims (10)

1. Use of an agent that inhibits the IL-34 signaling pathway in the manufacture of a medicament for the treatment of psoriasis.

2. Use of an agent that inhibits the IL-34 signaling pathway in the manufacture of a medicament for alleviating the progression of psoriasis lesions.

3. The use according to claim 2, wherein said alleviation of psoriatic lesion progression is achieved by inhibition of inflammatory cytosis, inhibition of keratinocyte proliferation and alleviation of pruritic symptoms.

4. The use according to any one of claims 1 to 3, wherein the medicament comprises a pharmaceutically acceptable adjuvant in addition to the agent which inhibits the IL-34 signalling pathway.

5. The use according to claim 4, wherein the pharmaceutical dosage form comprises an injection, a microneedle.

6. The use of claim 4, wherein the agent that inhibits the IL-34 signaling pathway is an agent that targets inhibition of IL-34 function.

7. The use of claim 6, wherein the agent that inhibits the IL-34 signaling pathway is an IL-34 antibody.

8. The use according to claim 7, wherein the IL-34 antibody is used in an amount of at least 5 μg/kg.

9. A medicament for treating psoriasis, which is characterized by comprising a preparation for inhibiting an IL-34 signal pathway and pharmaceutically acceptable auxiliary materials thereof.

10. The medicament of claim 9, wherein the agent that inhibits the IL-34 signaling pathway is an IL-34 antibody.

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