Use of GPR132 inhibitor for blocking CCL20-CCR6 axis in treatment and prevention of recurrence of psoriasis

Abstract

Provided in the present invention is the use of a GPR132 inhibitor for blocking the CCL20-CCR6 axis in the treatment and prevention of the recurrence of psoriasis. Specifically, provided in the present invention is the use of a GPR132 inhibitor for preparing a preparation or composition, wherein the preparation or composition is used for treating psoriasis in a subject in need thereof and / or preventing the recurrence of psoriasis in a subject in need thereof. Further provided in the present invention is a pharmaceutical product for treating psoriasis and / or preventing the recurrence of psoriasis. The pharmaceutical product provided in the present invention can be used to effectively treat, prevent or alleviate psoriasis and prevent the recurrence of psoriasis.

Description

Application of GPR132 inhibitor in blocking CCL20-CCR6 axis in treatment and prevention of recurrence of psoriasis TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, and more particularly to an application of a membrane receptor GPR132 inhibitor in treatment and prevention of recurrence of psoriasis. BACKGROUND

[0002] Psoriasis is a representative of inflammatory skin diseases and systemic inflammatory diseases, and is characterized by long course and repeated attacks. Even though a single antibody against IL-17A, a key pathogenic cytokine in the occurrence of psoriasis, has achieved significant therapeutic effect, there is still a problem of recurrence after drug withdrawal or even when the treatment has achieved significant therapeutic effect without drug withdrawal.

[0003] CCL20 is expressed in various tissues including skin, and plays an important role in immune homeostasis by binding to CCR6 without direct activation of the immune system. Since CCR6 is mainly expressed on IL-17-producing cells and is the only receptor for CCL20, the interaction between CCL20 and CCR6+ cells is called CCL20-CCR6 axis. Studies have shown that the CCL20-CCR6 axis plays a key role in the occurrence and recurrence of psoriasis. In normal human skin, CCL20 is expressed at very low concentrations in the epidermis. However, once CCL20 expression increases, CCR6+ cells are recruited to the CCL20 target, increasing the production of IL-17A, thereby exacerbating psoriasis.

[0004] Therefore, there is an urgent need in the art to develop a method for treating psoriasis and preventing its recurrence by blocking the CCL20-CCR6 axis. SUMMARY

[0005] The purpose of the present application is to provide an application of a GPR132 inhibitor in blocking CCL20-CCR6 axis in treatment and prevention of recurrence of psoriasis.

[0006] In a first aspect of the present application, there is provided a use of a G protein-coupled receptor 132 (GPR132) inhibitor for the manufacture of a preparation or a composition for treating psoriasis in a subject in need thereof and / or preventing recurrence of psoriasis in a subject in need thereof.

[0007] In another preferred embodiment, the subject in need thereof is a mammal.

[0008] In another preferred embodiment, the mammal includes a human or a non-human mammal.

[0009] In another preferred embodiment, the non-human mammal includes a rodent (such as a rat or a mouse), a primate (such as a monkey).

[0010] In another preferred embodiment, the desired subject is a patient with psoriasis.

[0011] In another preferred embodiment, the desired subject is a patient who has experienced a relapse of psoriasis.

[0012] In another preferred embodiment, the GPR132 inhibitor is selected from the group consisting of an antibody or a small molecule inhibitor targeting the GPR132 protein, a GPR132 gene targeting nucleic acid molecule or a gene editor, or a combination thereof.

[0013] In another preferred embodiment, the GPR132 gene targeting nucleic acid molecule comprises: an shRNA, an siRNA, an miRNA, or a combination thereof

[0014] In another preferred embodiment, the GPR132 gene targeting nucleic acid molecule comprises an antisense nucleic acid (such as an antisense oligonucleotide (ASO)).

[0015] In another preferred embodiment, the gene editor comprises a DNA gene editor and an RNA gene editor.

[0016] In another preferred embodiment, the gene editor comprises a gRNA and a gene editing protein.

[0017] In another preferred embodiment, the gRNA is an RNA that guides the gene editing protein to specifically bind to a gene corresponding to the GPR132 protein.

[0018] In another preferred embodiment, the gene editing protein is selected from the group consisting of CasRx, Cpf1, Cas9, Cas13a, Cas13b, Cas13c, or a combination thereof.

[0019] In another preferred embodiment, the GPR132 inhibitor is selected from the group consisting of GPR132-B-160, NOX-6-18, GPR132-siRNAs, GPR132 antibodies, or a combination thereof.

[0020] In a second aspect of the present application, there is provided a pharmaceutical product for treating psoriasis, the pharmaceutical product comprising:

[0021] (a) a first pharmaceutical composition comprising (a1) a GPR132 inhibitor, and (a2) a first pharmaceutically acceptable carrier; and

[0022] (b) a second pharmaceutical composition comprising (b1) another drug for treating psoriasis other than the GPR132 inhibitor or an active ingredient thereof, and (b2) a second pharmaceutically acceptable carrier.

[0023] In another preferred embodiment, the other drug for treating psoriasis or an active ingredient thereof other than the GPR132 inhibitor of (b1) is selected from the group consisting of a glucocorticoid, a LPC inhibitor, a CCL20 inhibitor, a CCR6 inhibitor, an IL-17A antagonist, a TNF-a antagonist, an IL-12 / IL-23 antagonist, a calcineurin inhibitor, a vitamin D3 derivative, or a combination thereof.

[0024] In another preferred embodiment, the other drug for treating psoriasis or an active ingredient thereof other than the GPR132 inhibitor of (b1) is selected from the group consisting of a LPC inhibitor, a CCL20 inhibitor, a CCR6 inhibitor, an IL-17A antagonist, or a combination thereof.

[0025] In another preferred embodiment, the other drug for treating psoriasis or an active ingredient thereof other than the GPR132 inhibitor of (b1) is selected from a LPC inhibitor.

[0026] In another preferred embodiment, the other drug for treating psoriasis or an active ingredient thereof other than the GPR132 inhibitor of (b1) is selected from an IL-17A antagonist.

[0027] In another preferred embodiment, the component (a1) in the first pharmaceutical composition accounts for 0.01-99.99 wt%, preferably 0.1-90 wt%, more preferably 1-80 wt% of the total weight of the first pharmaceutical composition.

[0028] In another preferred embodiment, the concentration of the component (a1) in the first pharmaceutical composition is 10 mg / ml-1000 mg / ml, preferably 20-500 mg / ml, more preferably 50-200 mg / ml.

[0029] In another preferred embodiment, the component (b1) in the second pharmaceutical composition accounts for 0.01-99.99 wt%, preferably 0.1-90 wt%, more preferably 1-80 wt% of the total weight of the second pharmaceutical composition.

[0030] In another preferred embodiment, the component (b1) in the second pharmaceutical composition is a LPC inhibitor, and (b1) accounts for 0.01-50 wt%, preferably 0.01-10 wt%, more preferably 0.1-5 wt% of the total weight of the second pharmaceutical composition.

[0031] In another preferred embodiment, the component (b1) in the second pharmaceutical composition is an IL-17A antagonist, and the concentration of (b1) in the second pharmaceutical composition is 1-1000 mg / ml, preferably 5-500 mg / ml, more preferably 10-100 mg / ml.

[0032] In another preferred embodiment, the first pharmaceutical composition is in a liquid, solid, or semi-solid form.

[0033] In another preferred embodiment, the dosage form of the first pharmaceutical composition is an oral dosage form, an injectable dosage form, or a topical pharmaceutical dosage form.

[0034] In another preferred embodiment, the dosage form of the first pharmaceutical composition includes a tablet, a granule, a capsule, an oral liquid, an injectable dosage form, a cream, an ointment, a gel, a spray, or a solution.

[0035] In another preferred embodiment, the carrier is selected from the group consisting of an infusion agent carrier and / or an injectable carrier; preferably, the carrier is one or more carriers selected from the group consisting of normal saline, dextrose saline, or a combination thereof.

[0036] In another preferred embodiment, the second pharmaceutical composition is in a liquid, solid, or semi-solid form.

[0037] In another preferred embodiment, the dosage form of the second pharmaceutical composition is an oral dosage form, an injectable dosage form, or a topical pharmaceutical dosage form.

[0038] In another preferred embodiment, the dosage form of the second pharmaceutical composition includes a tablet, a granule, a capsule, an oral liquid, an injectable dosage form, a cream, an ointment, a gel, a spray, or a solution.

[0039] In another preferred embodiment, the first pharmaceutical composition and the second pharmaceutical composition in the pharmaceutical product can be in the same dosage form or in different dosage forms.

[0040] In another preferred embodiment, the first pharmaceutical composition and the second pharmaceutical composition in the pharmaceutical product can be administered simultaneously or sequentially.

[0041] In a third aspect of the present application, there is provided a pharmaceutical product for preventing psoriasis relapse, the pharmaceutical product comprising:

[0042] (a) a first pharmaceutical composition comprising (a1) a GPR132 inhibitor, and (a2) a first pharmaceutically acceptable carrier; and

[0043] (b) a second pharmaceutical composition comprising (b1) another drug for preventing psoriasis relapse other than a GPR132 inhibitor or an active ingredient thereof, and (b2) a second pharmaceutically acceptable carrier.

[0044] In another preferred embodiment, the (b1) another drug for preventing psoriasis relapse other than a GPR132 inhibitor or an active ingredient thereof includes: a LPC inhibitor, a CCR6 inhibitor, or a combination thereof.

[0045] In another preferred embodiment, the (b1) other drug for preventing relapse of psoriasis or an active ingredient thereof other than the GPR132 inhibitor is selected from the group consisting of: an LPC inhibitor.

[0046] In another preferred embodiment, the (b1) other drug for preventing relapse of psoriasis or an active ingredient thereof other than the GPR132 inhibitor is selected from the group consisting of: a CCR6 inhibitor.

[0047] In another preferred embodiment, the (a1) in the first pharmaceutical composition is present in an amount of 0.01 to 99.99 wt%, preferably 0.1 to 90 wt%, more preferably 1 to 80 wt% based on the total weight of the first pharmaceutical composition.

[0048] In another preferred embodiment, the (a1) in the first pharmaceutical composition is present in an amount of 10 mg / ml to 1000 mg / ml, preferably 20 to 500 mg / ml, more preferably 50 to 200 mg / ml.

[0049] In another preferred embodiment, the (b1) in the second pharmaceutical composition is present in an amount of 0.01 to 99.99 wt%, preferably 0.1 to 90 wt%, more preferably 1 to 80 wt% based on the total weight of the second pharmaceutical composition.

[0050] In another preferred embodiment, the (b1) in the second pharmaceutical composition is an LPC inhibitor, and the (b1) is present in an amount of 0.01 to 50 wt%, preferably 0.01 to 10 wt%, more preferably 0.1 to 5 wt% based on the total weight of the second pharmaceutical composition.

[0051] In another preferred embodiment, the (b1) in the second pharmaceutical composition is a CCR6 inhibitor, and the (b1) is present in an amount of 1 to 1000 mg / ml, preferably 5 to 500 mg / ml, more preferably 10 to 100 mg / ml in the second pharmaceutical composition.

[0052] In another preferred embodiment, the first pharmaceutical composition is in a liquid, solid, or semi-solid form.

[0053] In another preferred embodiment, the first pharmaceutical composition is in a dosage form selected from the group consisting of: an oral dosage form, an injection, or a topical dosage form.

[0054] In another preferred embodiment, the first pharmaceutical composition is in a dosage form selected from the group consisting of: a tablet, a granule, a capsule, an oral solution, an injection, a cream, an ointment, a gel, a spray, or a solution.

[0055] In another preferred embodiment, the carrier is selected from the group consisting of: an infusion carrier and / or an injection carrier; preferably, the carrier is one or more carriers selected from the group consisting of: a physiological saline, a dextrose saline, or a combination thereof.

[0056] In another preferred embodiment, the second pharmaceutical composition is in a liquid, solid, or semi-solid form.

[0057] In another preferred embodiment, the dosage form of the second pharmaceutical composition is an oral dosage form, an injection, or a topical pharmaceutical dosage form.

[0058] In another preferred embodiment, the dosage form of the second pharmaceutical composition includes a tablet, a granule, a capsule, an oral liquid, an injection, a cream, an ointment, a gel, a spray, or a solution.

[0059] In another preferred embodiment, the first pharmaceutical composition and the second pharmaceutical composition in the pharmaceutical product can be prepared in the same dosage form or in different dosage forms.

[0060] In another preferred embodiment, the first pharmaceutical composition and the second pharmaceutical composition in the pharmaceutical product can be administered simultaneously or sequentially.

[0061] In a fourth aspect of the present application, there is provided a method for treating psoriasis, comprising the step of administering a GPR132 inhibitor or a pharmaceutical product as described in the second aspect of the present application to a subject in need thereof.

[0062] In a fifth aspect of the present application, there is provided a method for preventing the recurrence of psoriasis, comprising the step of administering a GPR132 inhibitor or a pharmaceutical product as described in the third aspect of the present application to a subject in need thereof.

[0063] In a sixth aspect of the present application, there is provided the use of a pharmaceutical product as described in the second aspect of the present application for the manufacture of a medicament for treating psoriasis.

[0064] In a seventh aspect of the present application, there is provided the use of a pharmaceutical product as described in the third aspect of the present application for the manufacture of a medicament for preventing the recurrence of psoriasis.

[0065] It should be understood that, within the scope of the present application, the above technical features of the present application and the technical features specifically described hereinafter (e.g. in the examples) can be combined with each other to form new or preferred technical solutions. Due to the limited space, they are not listed one by one here. BRIEF DESCRIPTION OF DRAWINGS

[0066] Figure 1 shows the increase of GPR132 expression and the endogenous agonist LPC content in the lesions of psoriasis patients at the onset and recurrence of the disease.

[0067] As shown in Figure 1A, the GPR132 expression in the non-lesion and lesion sites of psoriasis patients is significantly higher than that in healthy people;

[0068] As shown in FIG. IB, GPR132 expression in psoriasis patients' skin lesions was significantly increased after anti-IL-17A mAb treatment, while GPR132 expression in relapse was basically unchanged, and there was no significant difference in GPR132 expression after anti-IL-17A mAb treatment;

[0069] As shown in FIG. 1C and 1D, the content of GPR132 endogenous agonist LPC in psoriasis patients' skin lesions and serum was increased, although the content of LPC in their skin lesions and serum was decreased after anti-IL-17A mAb treatment, but increased again in relapse.

[0070] FIG. 2 shows that LPC aggravates psoriasis-like skin inflammation in mice.

[0071] As shown in FIG. 2A, mice with imiquimod (IMQ)-induced psoriasis showed obvious psoriasis-like inflammation, and the mice treated with IMQ and LPC showed more severe psoriasis-like inflammation, with reddish skin, more scales, and thicker epidermis. The results of H&E staining of the skin also confirmed the above results.

[0072] As shown in FIG. 2B, the proportion of CD45+ cells, neutrophils, γδ T cells, and IL-17A-producing γδ T cells in the skin of IMQ-treated psoriasis-like mice was significantly increased, and the proportion of the above cells in the skin of mice treated with IMQ and LPC was the highest, showing a further significant increase in cell proportion compared with IMQ-treated psoriasis-like mice.

[0073] FIG. 3 shows that LPC promotes keratinocyte secretion of CCL20 through GPR132.

[0074] As shown in FIG. 3A, the expression of CCL20 in the skin of mice treated with IMQ and LPC was up-regulated compared with IMQ-treated mice.

[0075] As shown in FIG. 3B, more CCR6+ γδ T cells were recruited in the skin of IMQ-treated psoriasis-like mice compared with control mice, and especially the CCR6+ γδ T cells recruited in the skin of mice treated with IMQ and LPC were the most, showing a further significant increase in CCR6+ γδ T cells compared with IMQ-treated psoriasis-like mice.

[0076] As shown in FIG. 3C and 3D, LPC promoted human and mouse keratinocytes to produce more CCL20, and the production of CCL20 in LPC-stimulated keratinocytes could be significantly inhibited by reducing the expression of GPR132 in human and mouse keratinocytes.

[0077] The above results show that LPC promotes keratinocyte secretion of CCL20 through GPR132, and the effect of LPC in promoting keratinocyte production of CCL20 can be significantly inhibited by reducing the expression of GPR132 in keratinocytes.

[0078] Figure 4 shows that LPC promotes CCL20 expression through GPR132 and recruits CCR6+ gd T cells, thereby exacerbating psoriatic skin inflammation.

[0079] As shown in Figure 4A, compared with WT mice (Gpr132 + / + mice), IMQ treatment or (IMQ+LPC) co-treatment of Gpr132 - / - mice reduced both erythema and scaling of the skin, and the epidermis was thinner, and even the dorsal skin of Gpr132 - / - mice treated with IMQ and LPC together did not develop typical psoriasis-like skin lesions;

[0080] As shown in Figure 4B, compared with WT mice (Gpr132 + / + mice), IMQ treatment of Gpr132 - / - mice significantly reduced the proportion of neutrophils (infiltration degree) in the skin, and the proportion of neutrophils (infiltration degree) in the skin of Gpr132 - / - mice treated with IMQ and LPC together also significantly decreased;

[0081] As shown in Figure 4C, compared with WT mice (Gpr132 + / + mice), IMQ treatment of Gpr132 - / - mice significantly reduced the expression of CCL20 in the skin, and the expression of CCL20 in the skin of Gpr132 - / - mice treated with IMQ and LPC together also significantly decreased;

[0082] As shown in Figure 4D, compared with WT mice (Gpr132 + / + mice), IMQ treatment or (IMQ+LPC) co-treatment of Gpr132 - / - mice significantly reduced the proportion of CCR6+ gd T cells in the skin, but the proportion of CCR6+ gd T cells in the lymph nodes of Gpr132 - / - mice treated with IMQ or (IMQ+LPC) co-treatment increased.

[0083] Figure 5 shows that LPC activates the CCL20 / CCR6 axis through GPR132 to participate in psoriasis recurrence.

[0084] As shown in Figure 5A, an IMQ-induced psoriasis-like mouse model was used to simulate the onset and recurrence time course of human psoriasis;

[0085] As shown in FIG. 5B, at the quiescent phase (resolution of inflammation), CCR6+ gd T cells were significantly increased in the skin of IMQ-treated or (IMQ+LPC) co-treated WT mice (Gpr132 + / + mice) compared to IMQ-untreated mice (Gpr132 + / + mice); while for Gpr132 - / - mice, even if they were IMQ-treated or (IMQ+LPC) co-treated, there was no significant difference in the CCR6+ gd T cells in their skin compared to IMQ-untreated WT mice (Gpr132 + / + mice);

[0086] In other words, the increase in CCR6+ gd T cells in the skin of mice caused by IMQ treatment or (IMQ+LPC) co-treatment was inhibited in Gpr132 - / - mice, and the CCR6+ gd T cells in the skin of IMQ-treated or (IMQ+LPC) co-treated Gpr132 - / - mice were significantly lower than those in IMQ-treated or (IMQ+LPC) co-treated WT mice (Gpr132 + / + mice);

[0087] Similarly, the CCR6+ gd T cells in the lymph nodes of IMQ-treated or (IMQ+LPC) co-treated WT mice were significantly increased compared to IMQ-untreated WT mice (Gpr132 + / + mice); while for Gpr132 - / - mice, even if they were IMQ-treated or (IMQ+LPC) co-treated, there was no significant difference in the CCR6+ gd T cells in their lymph nodes compared to IMQ-untreated WT mice (Gpr132 + / + mice) or even IMQ-untreated Gpr132 - / - mice, and the proportion of CCR6+ gd T cells was essentially unchanged;

[0088] In other words, the increase in CCR6+ gd T cells in the lymph nodes of mice caused by IMQ treatment or (IMQ+LPC) co-treatment was inhibited in Gpr132 - / - mice, and the CCR6+ gd T cells in the lymph nodes of IMQ-treated or (IMQ+LPC) co-treated Gpr132 - / - mice were significantly lower than those in IMQ-treated or (IMQ+LPC) co-treated WT mice (Gpr132 + / + mice);

[0089] As shown in FIG. 5C, when psoriasis was re-induced, the CCR6+ gd T cells in the skin of IMQ-treated or (IMQ+LPC) co-treated WT mice (Gpr132 + / +mice), psoriasiform lesions in the dorsal skin of WT mice previously treated with LPC and IMQ co-treatment were more severe; however, Gpr132 - / - mice showed only mild skin lesions when their dorsal skin was re-treated with IMQ; compared to WT mice (Gpr132 + / + mice), Gpr132 - / - mice showed significantly reduced skin lesions; the results were also confirmed by H&E staining of the skin;

[0090] As shown in Figure 5D, when psoriasis was re-induced, compared to WT mice previously treated with IMQ or (IMQ+LPC) co-treatment (Gpr132 + / + mice), Gpr132 - / - neutrophil infiltration in the mice was significantly reduced;

[0091] As shown in Figure 5E, when psoriasis was re-induced, compared to WT mice previously treated with IMQ or (IMQ+LPC) co-treatment (Gpr132 + / + mice), Gpr132 - / - CCL20 expression in the skin of the mice was significantly reduced;

[0092] In addition, when psoriasis was re-induced, compared to WT mice previously treated with IMQ only, CCL20 expression in the skin of WT mice previously treated with LPC and IMQ co-treatment was significantly increased, but Gpr132 - / - mice did not show significantly increased CCL20 expression in the skin;

[0093] As shown in Figure 5F, when psoriasis was re-induced, compared to WT mice previously treated with IMQ or (IMQ+LPC) co-treatment (Gpr132 + / + mice), Gpr132 - / - - the proportion of CCR6+ gd T cells in the skin of the mice was significantly reduced;

[0094] In addition, the proportion of CCR6+ gd T cells in the skin of WT mice previously co-treated with LPC and IMQ was significantly higher than that of WT mice previously treated with IMQ alone when psoriasis was induced again, while the proportion of CCR6+ gd T cells in the lymph nodes of WT mice previously co-treated with LPC and IMQ was significantly lower than that of WT mice previously treated with IMQ alone; while the proportion of CCR6+ gd T cells in the skin and lymph nodes of Gpr132 - / - In addition, the proportion of CCR6+ gd T cells in the skin of WT mice previously co-treated with LPC and IMQ was significantly higher than that of WT mice previously treated with IMQ alone when psoriasis was induced again, while the proportion of CCR6+ gd T cells in the lymph nodes of WT mice previously co-treated with LPC and IMQ was significantly lower than that of WT mice previously treated with IMQ alone; while the proportion of CCR6+ gd T cells in the skin and lymph nodes of Gpr132

[0095] Figure 6 shows that LPC promotes long-term changes in CCL20 through GPR132 and promotes psoriasis recurrence. Among them, Vehicle-Vas / Vas-vehicle is vaseline + solvent; Vehicle-IMQ is imiquimod + solvent; LPCs-IMQ is imiquimod + LPC.

[0096] As shown in Figure 6A, the experimental scheme of the LPC-induced mouse model is shown;

[0097] As shown in Figure 6B, the representative images of the back skin of WT mice and cKO mice in each treatment group and their H&E staining results are shown; + / + Mice and Gpr132 - / - Representative images of the back skin of mice and their H&E staining results;

[0098] As shown in Figure 6C, the representative images of the back skin of WT mice and cKO mice in each treatment group and their H&E staining results are shown; + / + Mice and Gpr132 - / - Representative flow cytometry images and statistical analysis of IL-17A+ gd T cells (IL-17A-producing gd T cells) and CCR6+ gd T lymphocytes in the skin of mice;

[0099] As shown in Figure 6D, the representative images of the back skin of WT mice and cKO mice in each treatment group and their H&E staining results are shown; + / + Mice and Gpr132 - / - Quantitative PCR analysis of CCL20 in the skin of mice.

[0100] Figure 7 shows that LPC activates keratinocytes (KCs) to produce CCL20 through GPR132 to promote psoriasis recurrence. Among them, Vehicle-Vas / Vas-vehicle is vaseline + solvent; Vehicle-IMQ is imiquimod + solvent; LPCs-IMQ is imiquimod + LPC.

[0101] As shown in Figure 7A, the experimental scheme of the LPC-induced mouse model is shown;

[0102] As shown in Figure 7B, the representative images of the back skin of WT mice and cKO mice in each treatment group and their H&E staining results are shown;

[0103] Representative flow cytometry plots and statistical analysis of IL-17A+ gd T cells (IL-17A-producing gd T cells) and CCR6+ gd T lymphocytes in the skin of WT mice and cKO mice in each treatment group are shown in FIG. 7C;

[0104] Quantitative PCR analysis of CCL20 in the skin of WT mice and cKO mice in each treatment group is shown in FIG. 7D. DETAILED DESCRIPTION

[0105] The inventors have made extensive and in-depth studies, and through a large number of screenings, for the first time, unexpectedly found a method for treating psoriasis and preventing psoriasis recurrence using a GPR132 inhibitor.

[0106] Specifically, the inventors found that the epidermis of mice treated with IMQ and LPC at the same time was thicker, showed more infiltration of neutrophils, and produced more pathogenic gd T17 cells by performing LPC on an IMQ-induced psoriasis mouse model. Transcriptomic analysis found that CCL20 expression in the skin of LPC-treated psoriasis mice was significantly increased. In vivo and in vitro experiments showed that LPC could induce keratinocytes to produce more CCL20 through GPR132, recruit more CCR6+ gd T17 cells, and amplify skin inflammation.

[0107] By simulating the process of drug withdrawal recurrence in clinical psoriasis patients, it was found that when the same part of the LPC-treated mice was again stimulated by IMQ, the expression of CCL20 in the skin was higher, more CCR6+ gd T17 cells were recruited, and psoriasis recurrence was accelerated or aggravated. Gpr132 - / - The mouse blocked the induction of CCL20 production by keratinocytes through GPR132 by LPC, which directly and effectively reduced CCR6+ gd T17 cells in the skin and lymph nodes, and effectively inhibited the migration of pathogenic CCR6+ gd T cells from the lymph nodes to the skin during psoriasis recurrence, so inhibiting GPR132 expression could both directly alleviate the symptoms of psoriasis and prevent psoriasis recurrence.

[0108] Especially for psoriasis patients treated with anti-IL17A mAb, the expression of GPR132 and / or LPC often has a significantly increased expression level after anti-IL17A mAb treatment or when psoriasis recurs. The administration of a GPR132 inhibitor to such patients to inhibit GPR132 expression can effectively block the role of LPC in promoting the CCL20-CCR6 axis through GPR132, thereby effectively preventing the recurrence of psoriasis in such patients. On this basis, the present application is completed.

[0109] TERMS

[0110] To facilitate a better understanding of the present disclosure, certain terms are defined first. As used in this application, and unless specifically identified otherwise, each of the following terms shall have the meaning given below.

[0111] The term“about” can refer to a value or composition that is within an acceptable error range for the particular value or composition as determined by one of ordinary skill in the art, which will depend in part on how the value or composition is measured or determined.

[0112] The term“administering” refers to physically introducing the product of the application into a subject using any of a variety of methods and delivery systems known to those in the art, including intravenous, intratumoral, intramuscular, subcutaneous, intraperitoneal, spinal, topical, or other parenteral routes of administration, such as by injection or infusion.

[0113] GPR132 / GPCR132

[0114] As used herein, the terms“GPR132”,“GPCR132” are used interchangeably, and all refer to G protein-coupled receptor 132, Gene ID: 29933 for human GPR132, Gene ID: 56696 for murine GPR132.

[0115] G protein-coupled receptor 132 (GPR132 / GPCR132) is one of the members of the G protein-coupled receptor (GPCR) superfamily. G protein-coupled receptors are cell surface receptors that couple to trimeric G proteins and have seven transmembrane alpha helices, and are the largest family of receptors discovered to date, with a large number of members. When a ligand binds to these receptors, it activates the coupled G protein, which in turn initiates different signal transduction pathways and leads to various biological effects. Known ligands that bind to G protein-coupled receptors include odors, pheromones, hormones, neurotransmitters, chemokines, and the like. These receptors can be small molecules such as sugars, lipids, polypeptides, or biological macromolecules such as proteins. Some special G protein-coupled receptors can also be activated by non-chemical stimuli, such as rhodopsin in photoreceptor cells which can be activated by light.

[0116] GPR132 antagonists / GPR132 inhibitors known in the prior art are described, for example, in GPR132-B-160 (CAS: 898211-21-7) with an EC50 value of 0.075 µM. In addition, GPR132-B-160 is also disclosed in patent application CN116903563A (Jia Ning et al., Application No. 202310421384.3, Patent Title: “GPCR modulators and uses thereof”), which has an activity of promoting insulin secretion with an EC50 value of 0.7 µM.

[0117] NOX-6-18 was used as a GPR132 inhibitor to reduce plasma glucose levels in high-fat diet-fed mice in the article Functional screening and rational design of compounds targeting GPR132 to treat diabetes, with insulin secretion-promoting activity.

[0118] lysophosphatidylcholine (LPC)

[0119] Lysophosphatidylcholine (LPC) is one of the main components of oxidized low-density lipoprotein (oxLDL). LPC containing saturated acyl chains is an important component of cell membranes and can be produced by the hydrolysis of phosphatidylcholine (PC) under the action of phospholipase A2 (PLA2). LPC containing saturated acyl chains can also be produced by transferring fatty acids to free cholesterol by lecithin-cholesterol acyltransferase (LCAT). A variety of LPC containing unsaturated acyl chains can be produced under the action of endothelial lipase (EL) and hepatic lipase (HL). In addition, under the action of acyl-CoA, LPC can be converted to PC again by lysophosphatidylcholine acyltransferase (LPCAT). LPC is degraded under the catalysis of lysophospholipase A1, C and D, among which autotaxin (ATX) is a protein with lysophospholipase D activity that can degrade LPC to produce lysophosphatidic acid (LPA), a lipid highly associated with skin diseases and cancer.

[0120] Although the human body contains high concentrations of LPC as high as 100 μM, the concentration and role of LPC in different disease mechanisms are different. As a secondary messenger, high concentrations of LPC play an important role in a variety of inflammatory diseases such as diabetes, atherosclerosis, and autoimmune diseases. The content of LPC with different acyl chains in the skin lesions and serum of psoriasis patients is significantly increased. Subcutaneous injection of LPC can induce skin T cell, monocyte and neutrophil infiltration to promote the development of skin inflammation. However, the specific role of LPC in the pathogenesis and recurrence mechanism of psoriasis still needs further study.

[0121] Psoriasis occurrence and recurrence

[0122] The pathogenesis of psoriasis is very complex, involving genetic predisposition, environmental triggers, disruption of skin barrier function, and dysfunction of the immune system. Different cells are involved in different stages of disease occurrence, development, stability, recurrence, etc. The main immunological event of psoriasis occurrence is the activation of skin keratinocytes by inflammatory factors such as TNF-a, IL-23 and IL17, and it has been reported that IL-17 is a key pathogenic product of psoriasis occurrence. From this, the method of targeting cytokines to treat psoriasis has emerged.

[0123] Although biological agents are a major innovation in the treatment of psoriasis, a considerable proportion of patients gradually lose efficacy and lead to disease recurrence as the treatment time is prolonged after initial use of biological agents to achieve disease remission, and some even progress to more serious types of psoriasis such as pustular, erythrodermic and arthropathic psoriasis. Among the above numerous therapeutic drugs, the history of topical glucocorticoids is the longest, and they are the most widely used. The recurrence after discontinuation of glucocorticoids and the recurrence and aggravation during continuous use are more distressing for both doctors and patients. Therefore, for the treatment of psoriasis, whether it is a traditional external preparation such as glucocorticoids or a new biological agent, the efficacy will decrease as the treatment time is prolonged, and most patients will experience a short remission period, and the disease will relapse after a few weeks or months, or even be more severe than before.

[0124] Regarding the mechanism of production of pathogenic product IL-17: As described above, CCL20 is expressed in various tissues including the skin, and plays an important role in immune homeostasis by binding to CCR6 without direct activation of the immune system. Given that CCR6 is mainly expressed on IL-17-producing cells and is the only receptor for CCL20, the interaction between CCL20 and CCR6+ cells is referred to as the CCL20-CCR6 axis. Studies have shown that the CCL20-CCR6 axis plays a key role in the occurrence and recurrence of psoriasis. In normal human skin, CCL20 is expressed at very low concentrations in the epidermis. However, once CCL20 expression increases, CCR6+ cells are recruited to the CCL20 target, increasing the production of IL-17A and thus exacerbating psoriasis.

[0125] Further, according to the previous research of the present inventors, the key mechanism of psoriasis recurrence is the migration / re-distribution of pathogenic CCR6+ γδT17 cells (CCR6+ γδT cells producing IL-17) between the skin and lymph nodes. During the quiescent phase of psoriasis (inflammation subsides), CCR6+ γδT17 cells in the skin migrate to the lymph nodes; and when psoriasis recurs, CCR6+ γδT17 cells in the lymph nodes migrate from the lymph nodes back to the skin, accelerating or exacerbating the recurrence of psoriasis.

[0126] In the present application, it is found for the first time that by inhibiting the expression of GPR132, on the one hand, the aggregation of pathogenic CCR6+ γδT cells in the skin and lymph nodes can be effectively reduced directly, and on the other hand, the migration of pathogenic CCR6+ γδT cells from the lymph nodes to the skin during the recurrence of psoriasis can be inhibited, so that inhibiting the expression of GPR132 can not only alleviate the symptoms of psoriasis, but also prevent the recurrence of psoriasis.

[0127] In addition, for psoriasis patients with elevated GPR132 expression and / or elevated lysophosphatidylcholine (LPC), or psoriasis relapse patients (such patients are usually patients after anti-IL17A monoclonal antibody treatment), by inhibiting GPR132 expression, the effect of LPC in promoting keratinocyte production of CCL20 can be further effectively blocked, the CCL20-CCR6 axis is blocked, the recruitment of CCL20 to CCR6+ gd T cells is inhibited, the aggregation of psoriasis pathogenic CCR6+ gd T cells in the skin is reduced, thereby reducing psoriasis symptoms and preventing psoriasis relapse.

[0128] Main advantages of the present application

[0129] 1. The present application first found that by inhibiting GPR132 activation in the skin to reduce the expression of CCL20, the recruitment of pathogenic CCR6+ gd T17 cells can be reduced, thereby treating psoriasis;

[0130] 2. The present application can inhibit the redistribution of CCR6+ gd T17 cells in lymph nodes and skin by inhibiting GPR132 activation in the skin to reduce the expression of CCL20, thereby effectively preventing or alleviating psoriasis relapse;

[0131] 3. Compared with general drugs for treating psoriasis, the present application can not only treat psoriasis but also prevent psoriasis relapse.

[0132] 4. For psoriasis patients after anti-IL17A monoclonal antibody treatment, further administration of a GPR132 inhibitor can effectively prevent the relapse of their psoriasis.

[0133] The present application will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present application and not used to limit the scope of the present application. The experimental methods in the following examples, if not specified, are generally carried out according to conventional conditions, for example, the conditions described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are weight percentages and weight parts.

[0134] Example 1: GPR132 expression and LPC content in skin lesions of psoriasis patients during onset and relapse are increased

[0135] In this example, as shown in FIG. 1A, it was found by analyzing public databases (GSE13355) that the expression of GPR132 in the skin lesions and non-lesion sites of psoriasis patients was significantly increased compared with healthy people.

[0136] Recruited psoriasis vulgaris patients were treated with anti-IL-17A mAb for 4 weeks, then drug withdrawal, and waiting for disease relapse. The skin lesions of these patients were collected before and after anti-IL-17A mAb treatment and after disease relapse for RNA-seq. As shown in FIG. IB, the results found that the expression of GPR132 in the skin of patients after anti-IL-17A mAb treatment was increased compared with that before anti-IL-17A mAb treatment, and the expression of GPR132 at relapse was not significantly reduced compared with that after anti-IL-17A mAb treatment.

[0137] Further comparison of the metabolome of skin lesions and serum before and after anti-IL-17A mAb treatment and after disease relapse, the results found that, as shown in FIG. 1C and FIG. ID, the content of GPR132 endogenous agonist lysophosphatidylcholine (LPC) in the skin lesions and serum of psoriasis patients was increased, although the LPC decreased after anti-IL-17A mAb treatment, but the LPC increased again at relapse.

[0138] Example 2: LPC aggravates psoriasiform skin inflammation in mice

[0139] In this example, a psoriasis mouse model was constructed by applying imiquimod (IMQ). The effect of skin GPR132 activation on the pathogenesis of psoriasis was explored by subcutaneous injection of saline solution containing or not containing LPC (‘solvent’ in FIG. 2, i.e. saline solution not containing LPC, ‘LPC’ in FIG. 2, i.e. saline solution containing LPC).

[0140] As shown in FIG. 2A, imiquimod (IMQ)-induced psoriasis model mice showed obvious psoriasiform inflammation, and the psoriasis lesions of mice co-treated with IMQ and LPC were more severe, mainly manifested as more scales on the surface of the lesions, more redness of the skin, and thicker epidermis. The results of skin H&E staining also confirmed the above results.

[0141] As shown in FIG. 2B, the results of flow cytometry analysis showed that the proportion (infiltration degree) of CD45+ cells, neutrophils, γδ T cells, and IL-17A-producing γδ T cells in the skin of IMQ-treated psoriasiform mice was significantly increased, and the proportion of the above cells in the skin of mice co-treated with IMQ and LPC was the highest, showing a further significantly increased cell proportion compared with IMQ-treated psoriasiform mice.

[0142] Example 3: LPC promotes keratinocyte secretion of CCL20 through GPR132

[0143] In this embodiment, as shown in Figure 3A, through RNA-seq analysis of the skin tissue of psoriasis mouse models with or without LPC treatment, it was found that, compared with IMQ-treated mice, the CCL20 in the top 20 up-regulated genes in the skin tissue of mice treated with IMQ and LPC was significantly changed, and CCL20 was significantly up-regulated.

[0144] As shown in Figure 3B, flow cytometry analysis showed that, compared with control mice, more CCR6+ gd T cells were recruited in the skin of IMQ-treated psoriasis-like mice, especially in the skin of mice treated with IMQ and LPC, which showed a further significant increase in CCR6+ gd T cells compared with psoriasis-like mice treated with only IMQ.

[0145] As shown in Figures 3C and 3D, in vitro cell experiments found that LPC could promote the production of CCL20 by human and mouse keratinocytes through GPR132, and reducing the expression of GPR132 in keratinocytes could significantly inhibit the production of CCL20 induced by LPC.

[0146] The above results show that LPC promotes the secretion of CCL20 by keratinocytes through GPR132, and reducing the expression of GPR132 in keratinocytes can significantly inhibit the role of LPC in promoting the production of CCL20 by keratinocytes.

[0147] Example 4: LPC promotes the expression of CCL20 to recruit CCR6+ gd T cells through GPR132

[0148] In this embodiment, wild-type mice (WT, Gpr132 + / + ) and Gpr132 - / - mice were used to construct psoriasis-like mouse models by applying IMQ.

[0149] As shown in Figures 4A and 4B, compared with IMQ-treated WT mice (Gpr132 + / + mice), the skin erythema and scales of IMQ-treated Gpr132 - / - mice were reduced, and the neutrophil infiltration was reduced. The results of skin H&E staining also confirmed that the epidermis of IMQ-treated Gpr132 - / - mice was not thickened. Moreover, compared with IMQ and LPC co-treated WT mice, even the dorsal skin of IMQ and LPC co-treated Gpr132 - / - mice did not appear typical psoriasis-like skin lesions, and the epidermis of IMQ and LPC co-treated Gpr132 - / The neutrophil infiltration in the skin of mice was also significantly reduced.

[0150] As shown in FIG. 4C, the proportion of CCR6+ gd T cells in the skin of IMQ-treated Gpr132 - / - The expression of CCL20 in the skin of mice was significantly reduced, and the proportion of CCR6+ gd T cells in the skin of mice treated with IMQ and LPC was significantly reduced. - / - The expression of CCL20 in the skin of mice was also significantly reduced.

[0151] As shown in FIG. 4D, the proportion of CCR6+ gd T cells in the skin of IMQ-treated Gpr132 - / - The proportion of CCR6+ gd T cells in the skin of mice treated with IMQ and LPC was significantly reduced. - / - The proportion of CCR6+ gd T cells in the skin of mice was significantly reduced; but the proportion of CCR6+ gd T cells in the skin of IMQ-treated Gpr132 - / - The proportion of CCR6+ gd T cells in the skin of mice treated with IMQ and LPC was significantly reduced. - / - The proportion of CCR6+ gd T cells in the lymph nodes of mice was increased.

[0152] The above results show that after LPC promotes keratinocytes to secrete more CCL20 through GPR132, CCL20 recruits more psoriasis pathogenic CCR6+ gd T cells to accumulate in the skin, and by reducing the expression of GPR132 in keratinocytes, the expression of CCL20 can be significantly inhibited, the recruitment of CCR6+ gd T cells by CCL20 can be inhibited, and the accumulation of psoriasis pathogenic CCR6+ gd T cells in the skin can be reduced, thereby reducing the symptoms of psoriasis.

[0153] Example 5: LPC activates CCL20-CCR6 axis through GPR132 to participate in psoriasis recurrence

[0154] In this embodiment, an IMQ-induced psoriasis-like mouse model was used to simulate the pathogenesis and recurrence schedule of human psoriasis.

[0155] As shown in FIG. 5A, an IMQ-induced psoriasis-like mouse model was used to simulate the pathogenesis and recurrence schedule of human psoriasis, in which WT mice (Gpr132 + / + mice) and Gpr132 - / - mice were treated with IMQ and subcutaneously injected with saline solution containing or not containing LPC, treated for 5 days, then rested for 21 days, and then re-challenged with the same dose of IMQ for 5 days.

[0156] As shown in FIG. 5B, during the resting period (inflammation subsides), the proportion of CCR6+ gd T cells in the skin of IMQ-treated or (IMQ+LPC) co-treated WT mice (Gpr132 + / + mice) was significantly increased compared to mice without IMQ treatment (Gpr132 + / + mice); while for Gpr132 - / -Mice, even if treated with IMQ or co-treated with (IMQ+LPC), did not have a significant difference in the proportion of CCR6+ gd T cells in their skin compared to WT mice (Gpr132 + / + Mice) that were not treated with IMQ.

[0157] In other words, the increase in CCR6+ gd T cells in the skin of mice caused by treatment with IMQ or co-treatment with (IMQ+LPC) was inhibited in Gpr132 - / - Mice, which had significantly lower CCR6+ gd T cells in their skin compared to WT mice (Gpr132 - / - Mice) treated with IMQ or co-treated with (IMQ+LPC). + / + Mice.

[0158] Similarly, the proportion of CCR6+ gd T cells in the lymph nodes of WT mice treated with IMQ or co-treated with (IMQ+LPC) was significantly increased compared to WT mice (Gpr132 + / + Mice) that were not treated with IMQ. - / - Mice, even if treated with IMQ or co-treated with (IMQ+LPC), did not have a significant difference in the proportion of CCR6+ gd T cells in their lymph nodes compared to WT mice (Gpr132 + / + Mice) that were not treated with IMQ, or even Gpr132 - / - Mice that were not treated with IMQ, the proportion of CCR6+ gd T cells was essentially unchanged.

[0159] In other words, the increase in CCR6+ gd T cells in the lymph nodes of mice caused by treatment with IMQ or co-treatment with (IMQ+LPC) was inhibited in Gpr132 - / - Mice, which had significantly lower CCR6+ gd T cells in their lymph nodes compared to WT mice (Gpr132 - / - Mice) treated with IMQ or co-treated with (IMQ+LPC). + / + Mice.

[0160] As shown in Figure 5C, when psoriasis was re-induced, the psoriatic-like lesions in the dorsal skin of WT mice (Gpr132 + / + Mice) that had previously been co-treated with LPC and IMQ were more severe compared to WT mice that had previously been treated with IMQ only; however, Gpr132 - / - Mice, whether or not they had previously been treated with IMQ or co-treated with (IMQ+LPC), only exhibited mild skin lesions when their dorsal skin was re-treated with IMQ. Compared to WT mice (Gpr132 + / +mouse), Gpr132 - / - Mouse skin lesions were significantly reduced. Skin H&E staining results also confirmed the above results.

[0161] As shown in Figure 5D, upon re-induction of psoriasis, the proportion of CCR6+ gd T cells in the skin of WT mice previously treated with IMQ or co-treated with IMQ and LPC was significantly higher than that of WT mice previously treated with IMQ or co-treated with IMQ and LPC (Gpr132 + / + mouse), Gpr132 - / - Mouse neutrophil infiltration was significantly reduced.

[0162] As shown in Figure 5E, upon re-induction of psoriasis, the proportion of CCR6+ gd T cells in the skin of WT mice previously treated with IMQ or co-treated with IMQ and LPC was significantly higher than that of WT mice previously treated with IMQ or co-treated with IMQ and LPC (Gpr132 + / + mouse), Gpr132 - / - The expression of CCL20 in mouse skin was significantly reduced.

[0163] In addition, upon re-induction of psoriasis, the expression of CCL20 in the skin of WT mice previously co-treated with LPC and IMQ was significantly higher than that of WT mice previously treated with IMQ alone, while the expression of CCL20 in the skin of Gpr132 - / - mouse, there was no significant increase in the expression of CCL20 in the skin upon re-stimulation with IMQ.

[0164] As shown in Figure 5F, upon re-induction of psoriasis, the proportion of CCR6+ gd T cells in the skin of WT mice previously treated with IMQ or co-treated with IMQ and LPC was significantly higher than that of WT mice previously treated with IMQ or co-treated with IMQ and LPC (Gpr132 + / + mouse), Gpr132 - / - The proportion of CCR6+ gd T cells in mouse skin was significantly reduced;

[0165] In addition, upon re-induction of psoriasis, the proportion of CCR6+ gd T cells in the skin of WT mice previously co-treated with LPC and IMQ was significantly higher than that of WT mice previously treated with IMQ alone, while the proportion of CCR6+ gd T cells in the lymph nodes of WT mice previously co-treated with LPC and IMQ was significantly reduced; while the proportion of CCR6+ gd T cells in the skin and lymph nodes of Gpr132 - / - mouse, there was no significant change in the proportion of CCR6+ gd T cells in the skin and lymph nodes upon re-stimulation with IMQ.

[0166] The above results show that inhibiting GPR132 expression can not only effectively directly reduce the aggregation of pathogenic CCR6+ gd T cells in the skin and lymph nodes, but also inhibit the migration of pathogenic CCR6+ gd T cells from the lymph nodes to the skin during psoriasis recurrence, thus reducing the symptoms of psoriasis and preventing the recurrence of psoriasis.

[0167] In summary, inhibiting GPR132 expression will inhibit keratinocyte production of CCL20, reduce the recruitment of pathogenic CCR6+ gd T cells in the skin and the redistribution of pathogenic CCR6+ gd T cells during psoriasis recurrence, reduce psoriasis skin inflammation, and prevent or slow down psoriasis recurrence.

[0168] Example 6: Psoriasis is more severe in mice treated with LPC and then re-stimulated with IMQ

[0169] In this example, as shown in FIG. 6A, WT mice (Gpr132 + / + mice) and Gpr132 - / - mice were first treated with LPC, and then IMQ was applied to the mice after they were left to stand for 7 days to construct a psoriasis-like mouse model.

[0170] As shown in FIG. 6B, compared with WT mice that had not been previously treated with LPC, the psoriasis-like lesions on the back skin of WT mice that had been previously treated with LPC were more severe. However, for Gpr132 - / - mice that had been previously treated with LPC, only mild skin lesions were observed when the back skin was re-treated with IMQ, and the degree of skin lesions was significantly reduced. The results of skin H&E staining also confirmed the above results.

[0171] As shown in FIGS. 6C-6D, compared with WT mice that had not been previously treated with LPC, the expression of CCL20 and the proportion of IL-17A+ gd T cells and CCR6+ gd T17 cells in the back skin of WT mice that had been previously treated with LPC were significantly increased after re-stimulation with IMQ. However, for Gpr132 - / - mice that had been previously treated with LPC, the changes in the expression of CCL20 and the proportion of IL-17A+ gd T cells and CCR6+ gd T17 cells in the skin were not significant when re-stimulated with IMQ.

[0172] Example 7: LPC promotes psoriasis recurrence by activating keratinocytes to produce CCL20 through GPR132

[0173] In Example 3, it was found that LPC promotes keratinocytes to produce CCL20 through GPR132. In this example, a specific gene knockout mouse (cKO mouse) of keratinocytes with GPR132 knockout was constructed.

[0174] As shown in FIG. 7A, the WT mice and cKO mice were first treated with LPC, and then treated with IMQ after the mice were rested for 7 days to construct the psoriasis-like mouse model.

[0175] As shown in FIG. 7B, compared with the WT mice without LPC treatment, the WT mice with LPC treatment showed more severe psoriasis-like lesions on the back skin. However, the cKO mice with LPC treatment showed only mild skin lesions on the back skin after IMQ treatment, and the degree of skin lesions was significantly reduced. The results of skin H&E staining also confirmed the above results.

[0176] As shown in FIGS. 7C-7D, compared with the WT mice without LPC treatment, the WT mice with LPC treatment showed significantly increased expression of CCL20, proportion of IL-17A+ gd T cells, and proportion of CCR6+ gd T17 cells in the back skin after IMQ restimulation. However, for the cKO mice with LPC treatment, the changes in the expression of CCL20, proportion of IL-17A+ gd T cells, and proportion of CCR6+ gd T17 cells in the skin were not significant after IMQ restimulation.

[0177] Example 8: GPR132 inhibitor / blocker reduces and slows down the recurrence of psoriasis skin

[0178] In this example, the IMQ was used to construct the psoriasis-like mouse model. By subcutaneously injecting normal saline with or without GPR132 inhibitor / blocker (GPR132-B-160, CAS No. 898211-21-7) at the same time of the first IMQ stimulation, the effect of GPR132 inhibitor / blocker on the onset of psoriasis was explored.

[0179] The results showed that the psoriasis lesions of the mice treated with IMQ and GPR132 inhibitor / blocker were significantly reduced, and the response rate reached 90%, compared with the mice treated with IMQ without GPR132 inhibitor / blocker treatment.

[0180] In addition, the IMQ-induced psoriasis-like mouse model was also used in this example to simulate the pathogenesis and recurrence schedule of human psoriasis. After the mice rested for inflammation subsidence, when the IMQ was restimulated, by subcutaneously injecting normal saline with or without GPR132 inhibitor / blocker (GPR132-B-160, CAS No. 898211-21-7), the effect of GPR132 inhibitor / blocker on the recurrence of psoriasis was explored.

[0181] The results show that, compared with mice not treated with GPR132 inhibitors / blockers, mice treated with GPR132 inhibitors / blockers have significantly reduced skin inflammation upon IMQ re-stimulation, with a response rate of 75%.

[0182] Therefore, GPR132 inhibitors / blockers can significantly reduce skin lesions in 90% of mice upon the first IMQ stimulation, and significantly reduce skin inflammation; and can significantly reduce skin lesions in 75% of mice upon IMQ re-stimulation, and significantly reduce skin inflammation.

[0183] The above results show that GPR132 inhibitors / blockers can play a preventive and / or therapeutic role in the pathogenesis of psoriasis and in the recurrence of psoriasis. GPR132 inhibitors / blockers are effective preventive and / or therapeutic agents for the pathogenesis of psoriasis and the recurrence of psoriasis.

[0184] All documents mentioned in the present application are incorporated herein by reference as if each document were individually incorporated. In addition, it is to be understood that various alterations and modifications can be made to the present application upon reading and understanding the above lecture of the present application, and these equivalent forms also fall within the scope of the appended claims.

[0185] Discussion

[0186] The recurrence of psoriasis has always been a difficult problem in the treatment and cure of psoriasis. The inventors' previous studies have shown that the CCL20-CCR6 axis plays a key role in the recurrence of psoriasis, and more importantly, CCR6+ γδT17 cells participate in the recurrence of psoriasis by migrating between the skin and lymph nodes. Previous studies have found that cytokines released by inflammatory cells can induce keratinocytes to secrete CCL20, and blocking the effects of these inflammatory factors on keratinocytes can reduce CCL20 expression, block CCR6+ cell recruitment, and continuously reduce the inflammation and damage of skin lesions in psoriasis patients. Therefore, it is very meaningful to find stimulants that induce the production of CCL20 in order to determine new potential therapeutic targets for the treatment and prevention of the recurrence of psoriasis.

[0187] The inventors found that G protein-coupled receptor 132 (GPR132) expression in the skin lesions of psoriasis patients was increased, and the expression was even higher after treatment with anti-IL-17A monoclonal antibody, and the expression in the skin lesions did not change when the psoriasis patients relapsed. Metabolomics found that the endogenous agonist of GPR132, lysophosphatidylcholine (LPC), was significantly increased in the skin lesions and serum of psoriasis patients. After treatment with anti-IL-17A monoclonal antibody, the increased LPC content in the skin lesions and serum returned to the normal level, but the LPC content in the skin lesions and serum of the patients again increased when the patients relapsed. Correlation analysis showed that the LPC content in the skin lesions and serum of psoriasis patients had a significant correlation with IL-17A in the skin lesions.

[0188] Through experiments on mice with psoriasis induced by imiquimod (IMQ), it was found that LPC could induce keratinocytes to produce more CCL20 through GPR132, recruit more CCR6+γδT17 cells, cause and aggravate skin inflammation; in the resting period (inflammation subsides), CCR6+γδT17 cells in the skin migrate to the lymph nodes; when the same part of the mouse is stimulated again by IMQ, the skin of the mouse previously treated with LPC can recruit more CCR6+γδT17 cells from the lymph nodes to migrate back to the skin, accelerate or aggravate psoriasis recurrence. Gpr132 - / - Mice not only block the recruitment of CCR6+γδT17 by LPC through GPR132 to induce keratinocytes to produce CCL20, aggravate psoriasis skin inflammation, but also block the migration of CCR6+γδT17 between the skin and the lymph nodes, preventing psoriasis recurrence.

[0189] Therefore, it is found for the first time in the present application that by inhibiting the keratinocyte membrane receptor GPR132 to block CCL20 secretion, the recruitment of pathogenic CCR6+γδT17 cells in the skin can be reduced, so as to achieve the purpose of treating and preventing psoriasis recurrence.

Claims

1. The use of a G protein-coupled receptor 132 (GPR132) inhibitor, characterized in that, This preparation is used to develop a formulation or composition for treating psoriasis in a desired subject and / or preventing psoriasis recurrence in a desired subject.

2. The use as described in claim 1, characterized in that, The target group is patients with psoriasis.

3. The use as described in claim 1, characterized in that, The target group is patients who have experienced a relapse of psoriasis.

4. The use as described in claim 1, characterized in that, The GPR132 inhibitor is selected from the group consisting of antibodies or small molecule inhibitors targeting the GPR132 protein, GPR132 gene-targeting nucleic acid molecules or gene editors, or combinations thereof.

5. The use as described in claim 1, characterized in that, The GPR132 inhibitor is selected from: GPR132-B-160, NOX-6-18, GPR132-siRNAs, GPR132 antibodies, or combinations thereof.

6. A pharmaceutical product for treating psoriasis, characterized in that, The pharmaceutical products include: (a) a first pharmaceutical composition comprising (a1) a GPR132 inhibitor and (a2) a first pharmaceutically acceptable carrier; and (b) A second pharmaceutical composition comprising (b1) a drug or active ingredient thereof for the treatment of psoriasis other than a GPR132 inhibitor, and (b2) a second pharmaceutically acceptable carrier.

7. The pharmaceutical product as described in claim 6, characterized in that, Other drugs or active ingredients used to treat psoriasis besides GPR132 inhibitors include: glucocorticoids, LPC inhibitors, CCL20 inhibitors, CCR6 inhibitors, IL-17A antagonists, TNF-α antagonists, IL-12 / IL-23 antagonists, calcineurin inhibitors, vitamin D3 derivatives, or combinations thereof; Preferably, the other drugs or active ingredients for treating psoriasis besides GPR132 inhibitors (b1) are selected from the group consisting of LPC inhibitors, CCL20 inhibitors, CCR6 inhibitors, IL-17A antagonists, or combinations thereof.

8. The pharmaceutical product as described in claim 6, characterized in that, The dosage form of the first or second pharmaceutical composition includes tablets, granules, capsules, oral liquids, injections, creams, ointments, gels, sprays, or solutions.

9. A pharmaceutical product for preventing psoriasis recurrence, characterized in that, The pharmaceutical products include: (a) a first pharmaceutical composition comprising (a1) a GPR132 inhibitor and (a2) a first pharmaceutically acceptable carrier; and (b) A second pharmaceutical composition comprising (b1) a drug or active ingredient thereof for the prevention of psoriasis recurrence other than a GPR132 inhibitor, and (b2) a second pharmaceutically acceptable carrier.

10. The pharmaceutical product as described in claim 9, characterized in that, Other drugs or active ingredients used to prevent psoriasis recurrence besides GPR132 inhibitors include: LPC inhibitors, CCR6 inhibitors, or combinations thereof.

11. The use of a pharmaceutical product as described in any one of claims 6-8, characterized in that, Used to prepare drugs for treating psoriasis.

12. The use of the pharmaceutical product as described in claim 9 or 10, characterized in that, Used to prepare drugs for preventing psoriasis recurrence.

13. A method for treating psoriasis, characterized in that, The method includes the step of administering a GPR132 inhibitor or a pharmaceutical product as described in any one of claims 6-8 to the desired subject.

14. A method for preventing psoriasis recurrence, characterized in that, The method includes the step of administering a GPR132 inhibitor or the pharmaceutical product as described in claim 9 or 10 to the desired subject.