Application of an ATX inhibitor in the preparation of a drug for treating psoriasis and its pharmaceutical composition

Abstract

This invention provides a novel ATX inhibitor, 2-[4-(4-chlorophenyl)-1,3-thiazolyl-2-yl]-3-pyridin-3-propyl-2-enyl nitrile, and its application, specifically in the preparation of drugs for treating psoriasis. Animal experiments show that both oral and topical administration have significant therapeutic effects on psoriatic mice with minimal impact on body weight and no obvious toxic side effects. Furthermore, lysophospholipase D activity assays revealed that 2-[4-(4-chlorophenyl)-1,3-thiazolyl-2-yl]-3-pyridin-3-propyl-2-enyl nitrile effectively inhibits lysophospholipase D activity in cells. This invention provides a new direction for the treatment of psoriasis.

Description

Technical Field

[0001] This invention relates to the application of a novel ATX inhibitor in the preparation of drugs for treating psoriasis and its pharmaceutical composition, belonging to the field of medicine. Background Technology

[0002] Psoriasis is a chronic inflammatory disease characterized by scaly, erythematous plaques. It can occur anywhere on the body, but is most common on the scalp and extensor surfaces of the limbs, and symptoms often worsen in winter. Currently, the global prevalence of psoriasis is approximately 2-3%, and the World Health Organization reports that its incidence is increasing year by year. In the past 20 years, the incidence of psoriasis in China has increased nearly fourfold. The pathogenesis of psoriasis is very complex; genetics, immunity, and various environmental factors can all interact to induce its development. One of the pathophysiological characteristics of psoriasis is the abnormal proliferation and differentiation of keratinocytes (such as acanthosis pilaris and nucleus preservation within keratinocytes, or parakeratosis).

[0003] Eighty percent of patients with psoriasis vulgaris have mild cases, making topical treatment the preferred option. Currently, commonly used topical medications for psoriasis include corticosteroids, retinoic acid, vitamin D derivatives, anthraquinone, tar preparations, and calcineurin inhibitors. Anthraquinone, tar preparations, and corticosteroids have limited use due to significant side effects, while retinoic acid, calcineurin inhibitors, and vitamin D derivatives are not entirely effective. Recently marketed calcipotriol-betamethasone ointment, a combination of corticosteroids and vitamin D derivatives, is slightly more effective than calcipotriol, but it is expensive. The limited availability of topical medications and the tendency for tolerance to develop have long been bottlenecks in the treatment of psoriasis; therefore, the development of new drugs for treating psoriasis is of great significance.

[0004] ATX, or Autotaxin, is a secreted lysophosphatase D that hydrolyzes lysophosphatidylcholine (LPC) to produce lysophosphatidic acid (LPA). Autotaxin is secreted by different types of cells and exists in body fluids. It is the only member of the exonucleotide pyrophosphatase / phosphodiesterase family with lysophosphatase (lysoPLD) activity. L. Lei et al. disclosed that a novel ATX inhibitor, 8380, showed significant efficacy in treating a psoriasis-like mouse model (L. Lei et al., "Lysophosphatidic acid mediates the pathogenesis of psoriasis by activating keratinocytes through LPAR5," Signal Transduct Target Ther, vol.6, no.1, p.19, Jan 15 2021, doi:10.1038 / s41392-020-00379-1).

[0005] Discovering more effective treatments has become an urgent clinical problem. Summary of the Invention

[0006] To address the problems existing in the prior art, one of the objectives of this invention is to provide an ATX inhibitor, the structural formula of which is shown in Formula I:

[0007]

[0008] Accordingly, the chemical formula of the compound with structural formula I is C 17 H 10 ClN3S, chemically named 2-[4-(4-chlorophenyl)-1,3-thiazol-2-yl]-3-pyridin-3-ylprop-2-enenitrile, (Z)-2-[4-(4-chlorophenyl)-1,3-thiazol-2-yl]-3-pyridin-3-ylprop-2-enenitrile.

[0009] The second objective of this invention is to provide the use of the above-mentioned ATX inhibitor, specifically the use of 2-[4-(4-chlorophenyl)-1,3-thiazolyl-2-yl]-3-pyridin-3-ylprop-2-enonitrile in the preparation of a drug for treating psoriasis.

[0010] Preferably, the above-mentioned psoriasis is psoriasis vulgaris.

[0011] Preferably, the above-mentioned drug is a drug that alleviates and improves psoriasis by inhibiting the activity of lysophospholipase D in cells.

[0012] A third objective of this invention is to provide a pharmaceutical composition in which the aforementioned ATX inhibitor is the active ingredient.

[0013] Preferably, the above-mentioned drug is an oral drug; in a more preferred embodiment, the oral drug dose in a psoriasis-like mouse model is 20 mg / kg to 40 mg / kg.

[0014] Preferably, the above-mentioned drug is a topical drug; a more preferred embodiment is a topical cream drug.

[0015] Preferably, the topical medication further includes a topical matrix, and the topical medication is prepared by mixing the ATX inhibitor with the matrix.

[0016] More preferably, the matrix is ​​selected from white petrolatum, octadecanol, glyceryl monostearate, p-hydroxybenzoic acid, glycerol, sodium lauryl sulfate, and water; wherein white petrolatum, octadecanol, glyceryl monostearate, and p-hydroxybenzoic acid are the oil phase, and glycerol, sodium lauryl sulfate, and water are the aqueous phase.

[0017] The active ingredient in the topical medicine has a mass percentage of 0.5% to 1%, with an optimal value of 0.5%.

[0018] Compared with existing technologies, ATX inhibitors represent a novel therapeutic target. Animal experiments have shown that 2-[4-(4-chlorophenyl)-1,3-thiazolyl-2-yl]-3-pyridin-3-ylprop-2-enonitrile exhibits significant therapeutic effects in psoriatic mice, whether administered orally or topically, with minimal impact on body weight and no obvious toxic side effects observed. Furthermore, lysophospholipase D activity assays revealed that 2-[4-(4-chlorophenyl)-1,3-thiazolyl-2-yl]-3-pyridin-3-ylprop-2-enonitrile effectively inhibits lysophospholipase D activity in cells. Both systemic and topical administration demonstrate significant therapeutic effects, providing a new direction for the treatment of psoriasis. Attached Figure Description

[0019] Figure 1 This is a photograph of the mouse ear in an animal experiment using the oral formulation obtained in Example 1;

[0020] Figure 2 This is a diagram showing the HE staining results of the oral formulation obtained in Example 1 in animal experiments;

[0021] Figure 3 This is a graph showing the statistical results of animal experiments for the oral formulation obtained in Example 1;

[0022] Figure 4 This is a photograph of the mouse ear in an animal experiment using the oral formulation obtained in Example 1;

[0023] Figure 5 This is a diagram showing the HE staining results of the oral formulation obtained in Example 1 in animal experiments;

[0024] Figure 6 This is a graph showing the statistical results of animal experiments for the oral formulation obtained in Example 1;

[0025] Figure 7 This is a photograph of the back of a mouse used in an animal experiment to obtain the topical formulation obtained in Example 1.

[0026] Figure 8 This is an image showing the HE staining results of an animal experiment on the topical preparation obtained in Example 1;

[0027] Figure 9 This is a graph showing the statistical results of animal experiments on the topical preparation obtained in Example 1;

[0028] Figure 10 The figure shows the experimental results of lysophospholipase D activity in Example 1. Detailed Implementation

[0029] The present invention will be further described in detail and completely below with reference to embodiments. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0030] Unless otherwise specified, the experimental methods in the following examples are generally performed under conventional conditions, such as those described in *Pharmacology Laboratory Manual*, 3rd Edition, Science Press, 2002. The methods for imiquimod (IMQ) modeling and psoriasis treatment efficacy are based on published literature ("Lysophosphatidylcholine facilitates the pathogenesis of psoriasis through activating keratinocytes and T cell differentiation via glycolysis" in *Journal of the European Academy of Dermatology and Venereology*) or according to the manufacturer's recommendations. Unless otherwise specified, all reagents involved in the embodiments of this invention are commercially available products and can be purchased through commercial channels.

[0031] The main materials involved in the embodiments of this invention are as follows:

[0032] 1. Laboratory animals

[0033] Specific pathogen-free (SPF) grade BAL B / C male mice, 6-8 weeks old and weighing 20-23g, were purchased from Hunan Slack Jingda Laboratory Animal Co., Ltd. and housed at the Laboratory Animal Center of Central South University. This study has passed the laboratory animal welfare ethics review of Central South University. The "3R" principle was strictly followed during the experiment to minimize animal suffering and discomfort.

[0034] 2. Experimental reagents

[0035]

[0036] Example 1

[0037] I. Preparation of topical cream formulations

[0038] Weigh 25 mg of 2-[4-(4-chlorophenyl)-1,3-thiazolyl-2-yl]-3-pyridin-3-ylprop-2-enonitrile (hereinafter referred to as AI-219 / 3) using weighing paper and dissolve it in 250 μL of DMSO to prepare a solution for later use. Add 0.4 g of octadecyl alcohol, 0.6 g of white petrolatum, 0.01 g of p-hydroxybenzoic acid, and 0.1 g of glyceryl monostearate to a beaker according to the instructions, and then place it in a preheated 70°C constant temperature magnetic stirrer and heat it in a 70°C water bath to dissolve, obtaining the oil phase of the cream; at the same time, add 0.05 g of sodium dodecyl sulfate, 0.35 g of glycerin, and 3.5 g of water to another beaker according to the proportion and heat to dissolve, obtaining the aqueous phase of the cream. Then slowly add the aqueous phase to the oil phase and stir for 10 minutes. Finally, slowly add the AI-219 / 3 solution and continue stirring for 10 minutes to obtain the above-mentioned topical ointment for treating psoriasis. Let it stand at room temperature and stir it quickly with a glass rod until it reaches a semi-solid state at room temperature. Then bottle it for later use.

[0039] II. Preparation of oral formulations

[0040] The oral preparation was a gavage solution, which was prepared by dissolving AI-219 / 3 in sodium carboxymethyl cellulose (CMC-NA) solution to form a suspension for later use. (Oral administration group A: 0.8 mg AI-219 / 3 dissolved in 200 μL CMC-NA) (Oral administration group B: 0.4 mg AI-219 / 3 dissolved in 100 μL CMC-NA). Each gavage administration was 100 μL per animal, once a day for group A and twice a day for group B.

[0041] Comparative Example 1

[0042] The difference between this comparative example and Example 1 is that AI-219 / 3 is not added as an active ingredient, while the amounts of excipients used in the cream are the same as in Example 1. White petrolatum, octadecanol, glyceryl monostearate, and p-hydroxybenzoic acid were added to a beaker in proportion and placed in a DF-101S thermostatically heated magnetic stirrer. The mixture was heated in a 70°C water bath to dissolve the AI-219 / 3, and stirred for 10 minutes to obtain the oil phase of the cream. Simultaneously, glycerin, sodium lauryl sulfate, and water were added to another beaker in proportion and heated to dissolve the AI-219 / 3, stirring for 5 minutes to obtain the aqueous phase of the cream. The aqueous phase was then slowly added to the oil phase and stirred for 10 minutes. The mixture was then allowed to stand at room temperature and rapidly stirred with a glass rod until it reached a semi-solid state at room temperature, thus obtaining a control cream formulation without AI-219 / 3.

[0043] The alleviating effect of imiquimod (IMQ)-induced psoriasis in mice.

[0044] 1. Imiquimod (IMQ) modeling

[0045] Imiquimod-induced mouse models effectively mimic human psoriasis symptoms, including skin thickening, abnormal keratinocyte-related proteins, inflammatory cell infiltration, and related inflammatory cytokines. Due to its simplicity, ease of operation, and stability, this model has become one of the most widely used psoriasis models in research.

[0046] 1) Dosing regimen

[0047] Eighteen SPF-grade BALB / c mice aged 6-8 weeks were randomly divided into three groups of six each. The backs of the mice were shaved in a 2cm x 2cm area. The treatments for each group were as follows:

[0048] experimental group Administration method Blank control group (Control) No action taken IMQ control group (topical application) Three hours after applying imiquimod to the back, apply the cream preparation obtained in Comparative Example 1. Topical application group Three hours after applying imiquimod to the back, apply the cream preparation obtained in Example 1.

[0049] Twenty-four SPF-grade C57 mice aged 6-8 weeks were randomly divided into four groups, with six mice in each group. The treatment methods for each group were as follows:

[0050]

[0051] After 6 consecutive days of drug application in each group, to evaluate the severity of skin inflammation in mice, the Psoriasis Skin Lesion Area and Severity Index (PASI) was used to assess the psoriasis-like symptoms on the skin lesions on the back and ears of each group of mice. The severity of erythema and scales was categorized as follows: 0 (none); 1 (mild); 2 (moderate); 3 (obvious); 4 (severe). Ear thickness was measured using calipers. Daily food intake, body weight, PASI score, and ear thickness were recorded. Mice were then weighed, photographed, and euthanized. Skin lesions on the back and ears were collected for HE staining to examine epidermal thickness and keratinocyte proliferation.

[0052] 2) Experimental Results

[0053] Figures 1-3The figures show a comparison of experimental results between the blank control group, the IMQ control group, and the oral administration group A; among them... Figure 1 The image shows the phenotypic effect of the oral administration group A. Compared to the blank control group, the positive control group showed significantly more erythema, thickening, and scaling on the ear lesions, demonstrating the successful induction of the psoriasis model after imiquimod application. Compared to the IMQ control group, the oral administration group A showed a significant reduction in erythema, infiltration, and thickening, indicating that the oral administration group A effectively treats psoriasis. Figure 1 As shown, the ear skin treated with the IMQ control group exhibited significant thickening, accompanied by excessive proliferation of keratinocytes and infiltration of immune cells, while the oral administration group A significantly inhibited IMQ-induced psoriatic skin inflammation. Figure 2 As shown, HE staining also revealed significant pathophysiological changes in the lesions of the IMQ control group: hyperkeratosis with parakeratosis, thickened stratum corneum, epidermal hyperplasia, inflammatory cell infiltration, and visible Munro microabscesses. Figure 3 As shown, the skin lesions in the oral administration group A were significantly improved. Statistical analysis also revealed that in the psoriasis-like mouse model, the erythema, scaling, and infiltration of the skin lesions in the oral administration group A were significantly improved, while the body weight did not change significantly compared to the matrix group. (*P<0.05,**P<0.01,***P<0.001,****P<0.0001).

[0054] Figures 4-6 The figures show a comparison of experimental results between the blank control group, the IMQ control group, and the oral administration group B; among which... Figure 4 The image shows the phenotypic effect of the oral administration group B. Compared to the blank control group, the positive control group showed significantly more erythema, thickening, and scaling on the ear lesions, demonstrating the successful induction of the psoriasis model after imiquimod application. Compared to the IMQ control group, the oral administration group B showed a significant reduction in erythema, infiltration, and thickening, indicating that the oral administration group B effectively treats psoriasis. Figure 4 As shown, the ear skin treated with the IMQ control group exhibited significant thickening, accompanied by excessive proliferation of keratinocytes and infiltration of immune cells, while the oral administration group B significantly inhibited IMQ-induced psoriatic skin inflammation. Figure 5 As shown, HE staining also revealed significant pathophysiological changes in the lesions of the IMQ control group: hyperkeratosis with parakeratosis, thickened stratum corneum, epidermal hyperplasia, inflammatory cell infiltration, and visible Munro microabscesses. Figure 6 As shown, the skin lesions in the oral administration group B were significantly improved. Statistical analysis also revealed that in the psoriasis-like mouse model, the skin lesions, erythema, scaling, and infiltration in the oral administration group B were significantly improved, while the body weight did not change significantly compared to the matrix group. (*P<0.05,**P<0.01,***P<0.001,****P<0.0001).

[0055] Based on the above results, the treatment effect of oral administration group B is better than that of oral administration group A. In other words, at the same dosage, twice-daily administration is more effective than once-daily administration.

[0056] Figures 7-9 These are comparative figures showing the experimental results of the blank control group, the IMQ control group, and the topical application group; among them... Figure 7 The image shows the phenotypic effect of the topical treatment group. Compared to the blank control group, the positive control group showed significantly more erythema, thickening, and scaling on the back lesions, demonstrating the successful induction of the psoriasis model after imiquimod application. Compared to the IMQ control group, the topical treatment group showed a significant reduction in erythema, infiltration, and thickening, indicating that the topical treatment group effectively treats psoriasis. Figure 7 As shown, the back skin treated with the IMQ control group exhibited significant thickening, accompanied by excessive proliferation of keratinocytes and infiltration of immune cells, while the topical administration group significantly inhibited IMQ-induced psoriatic skin inflammation. Figure 8 As shown, HE staining also revealed significant pathophysiological changes in the skin lesions of the IMQ control group: hyperkeratosis with parakeratosis, thickened stratum corneum, epidermal hyperplasia, inflammatory cell infiltration, and visible Munro microabscesses. In contrast, the skin lesions in the topical treatment group showed significant improvement. Figure 9 As shown, statistical analysis also revealed that in the psoriasis-like mouse model, the topical treatment group showed significant improvement in erythema, scaling, and infiltration of skin lesions, while the body weight did not change significantly compared to the matrix group. (*P<0.05,**P<0.01,***P<0.001,****P<0.0001).

[0057] Lysophospholipase D activity assay

[0058] (1) lyso-PLD assay buffer: Dissolve 1.02g MgCl2, 12.114g Tris and 29.22g NaCl in 800mL ddH2O, adjust the pH to 8.0, and bring the volume to 1000mL. Store at room temperature. (2) 10×PAT: Dissolve 5mg horseradish peroxidase in 17.72mL lyso-PLD assay buffer, then add 1.68mL 250mM 4-AAP and 0.6mL 500mM 3-(N-ethyl-m-toluidine)-2-hydroxypropanesulfonate sodium salt (TOOS). Store at -20℃. (3) After seeding HaCaT cells in 96-well plates overnight, add Hepes liquid (stimulated with IL22 and AI-219 / 3 10uM) for 24 hours. Take 20ul of Hepes liquid, add 138μL of buffer, 20μL of 1×PAT, 20μL of 10mM LPC, and 2μL of choline oxidase (100U / ml). Measure the absorbance at 550nm.

[0059] After seeding HaCaT cells in 96-well plates overnight, Hepes medium (stimulated with IL22 and AI-219 / 34612004 10 μM) was added for 24 hours. Then, 20 μL of Hepes medium was taken and mixed with 138 μL of buffer, 20 μL of 1×PAT, 20 μL of 10 mM LPC, and 2 μL of choline oxidase (100 U / ml). The absorbance at 550 nm was measured.

[0060] Assays showed that AI-219 / 3 effectively inhibited the activity of lysophospholipase D in cells. Figure 10 As shown, IL22 stimulation in HaCaT cells leads to an increase in lysophospholipase D activity. Then, after simultaneous treatment with IL22 and AI-219 / 3, it was found that lysophospholipase D activity was significantly reduced compared with IL22 stimulation alone, proving that AI-219 / 3, as an ATX inhibitor, can reduce lysophospholipase D activity.

[0061] Finally, it is necessary to state that the above embodiments are only used to further illustrate the technical solution of the present invention in detail, and should not be construed as limiting the scope of protection of the present invention. Any changes, modifications, substitutions, combinations, or simplifications made by those skilled in the art without departing from the scope of the technical solution of this application using the above-disclosed technical content should be considered as equivalent substitutions and are included within the scope of protection of the present invention.

Claims

1. The use of 2-[4-(4-chlorophenyl)-1,3-thiazolyl-2-yl]-3-pyridin-3-ylprop-2-enonitrile (the compound shown in Formula I) in the preparation of medicaments for treating psoriasis; ； I。 2. The application according to claim 1, characterized in that, The psoriasis mentioned is psoriasis vulgaris.

3. The application according to claim 1, characterized in that, The drug is an oral medication.

4. The application according to claim 1, characterized in that, The drug is for external use only.

5. The application according to claim 4, characterized in that, The topical medication also includes a topical matrix, which is prepared by mixing 2-[4-(4-chlorophenyl)-1,3-thiazolyl-2-yl]-3-pyridin-3-ylprop-2-enonitrile with the matrix.

6. The application according to claim 5, characterized in that, The active ingredient in the topical medication is 0.5% to 1% by mass.

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