Application of small molecule compound Z57346765 in preparation of medicine for treating psoriasis

Abstract

The invention belongs to the technical field of biological medicine, and discloses application of a small molecule compound Z57346765 and pharmaceutically acceptable salt thereof in preparation of a medicine for treating psoriasis. The Z57346765 and the pharmaceutically acceptable salt or the medicine thereof play a role in treating the psoriasis by inhibiting the expression or activity of the PGK1. The pharmaceutically acceptable carrier or excipient is selected from one or more of DMSO (Dimethylsulfoxide), PEG-300 (Polyethylene Glycol), Tween-80, normal saline, vegetable oil, mannitol, starch, lactose, magnesium stearate and hydroxypropyl methylcellulose.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to new uses of the small molecule compound Z57346765, particularly its application in the treatment of psoriasis, and psoriasis treatment drugs containing the compound. Background Technology

[0002] Psoriasis is a chronic, relapsing autoimmune skin disease with a complex pathogenesis closely related to multiple factors, including genetics, environment, immune disorders, and metabolic abnormalities. Clinically, it manifests as scaly erythematous patches or plaques on the skin. Pathological features include excessive proliferation and abnormal differentiation of keratinocytes, as well as inflammatory cell infiltration. In severe cases, it can affect the joints, cardiovascular system, and other systems, posing a significant threat to the patient's physical and mental health.

[0003] Currently, treatments for psoriasis mainly include topical medications (such as corticosteroids and vitamin D analogs), systemic medications (such as methotrexate), and biologics (such as anti-TNF-α and IL-17 antibodies). However, existing treatments have many shortcomings: long-term use of corticosteroids can easily lead to adverse reactions such as skin atrophy and pigmentation; biologics are expensive, and some patients have a poor response or risk of relapse; moreover, most treatments only relieve symptoms and cannot fundamentally intervene in the pathogenesis. Therefore, there is an important clinical need to develop novel therapeutic drugs with strong targeting, stable efficacy, and high safety.

[0004] PGK1 (phosphoglycerate kinase 1) is a key enzyme in the glycolysis pathway, responsible for catalyzing the conversion of 1,3-diphosphoglycerate to 3-phosphoglycerate, while simultaneously generating ATP, playing a central role in cellular energy metabolism. Existing research indicates that PGK1 is closely related to various diseases, promoting tumor cell proliferation and metastasis, and participating in the regulation of pathological processes in ischemic diseases. In the field of autoimmune diseases, metabolic reprogramming of immune cells (such as enhanced glycolysis) has been shown to be closely related to disease development and progression; however, current technology has not elucidated the specific expression characteristics and mechanism of action of PGK1 in psoriasis, nor has it found any inhibitors targeting PGK1 that can be used for the treatment of psoriasis.

[0005] Psoriasis treatment remains a global challenge, with existing drugs having many limitations, necessitating novel interventions to overcome clinical bottlenecks. Mild psoriasis is primarily treated with topical corticosteroids, calcipotriol, or a combination of both. However, potent corticosteroids are prone to adverse reactions such as subcutaneous fat atrophy and adrenal suppression, making long-term clinical use unsuitable. Oral medications such as methotrexate, cyclosporine, and retinoids are suitable for moderate to severe psoriasis that is difficult to control with topical medications, but they also have significant adverse reactions: methotrexate has significant hepatotoxicity and severe gastrointestinal reactions; cyclosporine is effective for severe psoriasis, but is accompanied by severe nephrotoxicity, gastrointestinal discomfort, and hirsutism; acitretin has clear teratogenicity and is contraindicated in pregnant women and those planning pregnancy, and may also cause symptoms such as dry skin, dyslipidemia, and elevated transaminase levels. Targeted biologic therapy has brought breakthroughs in psoriasis treatment, with selectively targeting the IL-17, IL-23, and TNF-α pathways significantly revolutionizing treatment modalities. TNF-α inhibitors are effective, but some patients experience symptom exacerbation or new-onset paradoxical psoriasis after treatment. The IL-17 inhibitor bulimikokumab carries a higher risk of oral candidiasis. Non-biological small molecule drugs such as JAK / TYK2 inhibitors and PDE-4 inhibitors also have limitations, including high cost, poor accessibility, high treatment failure rates, and insufficient long-term safety data. These shortcomings have created a bottleneck in psoriasis treatment, with frequent adverse reactions and relapse after drug discontinuation becoming core clinical challenges. The close relationship between PGK1 and the pathogenesis of psoriasis provides crucial theoretical support for the development of novel treatment options.

[0006] PGK1, as a key rate-limiting enzyme in glycolysis, directly regulates cellular energy metabolism. In psoriasis patients, keratinocytes and immune cells exhibit hyperglycolysis (the Warburg effect), and this metabolic reprogramming promotes the production of pro-inflammatory factors, thereby exacerbating skin inflammation. Studies have confirmed that the accumulation of 3-phosphoglycerate (3-PG) catalyzed by PGK1 can activate the serine synthesis pathway and regulate core inflammatory signaling pathways such as TNF-α. In Th17 cells, high PGK1 expression desensitizes them to immune regulatory signals; silencing PGK1 can restore its function by upregulating FOXP3 and IL-10. The expression of PHGDH, the rate-limiting enzyme in serine synthesis, is downregulated in psoriatic lesions, while 3-PG (which depends on PGK1 for production) is a key precursor in serine synthesis, suggesting the important role of the PGK1-3-PG-PHGDH regulatory axis—inhibition of PHGDH promotes TNF-α secretion by keratinocytes, activating psoriasis-related pathological pathways. Mouse model studies have confirmed that targeting glycolysis can effectively reduce inflammation, and PGK1 inhibitors can block the glycolysis pathway, highlighting the creative value and clinical potential of novel treatments targeting PGK1.

[0007] Z57346765 is a novel PGK1-targeting small molecule compound discovered through a high-throughput screening platform and co-screened and validated with CHR-6494. This compound specifically binds to PGK1, significantly inhibiting its glycolytic metabolic enzyme activity. By regulating cellular metabolism, DNA replication, and the expression of cell cycle-related genes, it inhibits the proliferation of clear cell renal cell carcinoma (KIRC) cells, exhibiting significant anti-KIRC activity both in vitro and in vivo, with superior efficacy and safety compared to CHR-6494. Considering the core regulatory role of PGK1 in the pathogenesis of psoriasis, the specific targeting advantage of Z57346765 provides a novel potential therapeutic direction for psoriasis, supporting the inventiveness and application value of this patented intervention program.

[0008] Furthermore, on the one hand, there are differences in understanding among those skilled in the art; on the other hand, the applicant studied a large number of documents and patents when making this invention, but due to space limitations, not all details and contents were listed in detail. However, this does not mean that the present invention does not possess the features of these prior art. On the contrary, the present invention already possesses all the features of the prior art, and the applicant reserves the right to add relevant prior art to the background art. Summary of the Invention

[0009] To address the shortcomings of existing technologies, this invention provides the application of a small molecule compound Z57346765 and its pharmaceutically acceptable salt in the preparation of a drug for treating psoriasis, wherein the structural formula of the small molecule compound Z57346765 is: .

[0010] According to a preferred embodiment, Z57346765 and its pharmaceutically acceptable salts, or the drug, exert a psoriasis therapeutic effect by inhibiting the expression or activity of PGK1.

[0011] According to a preferred embodiment, the drug comprises an effective dose of the small molecule compound Z57346765 and a pharmaceutically acceptable carrier or excipient.

[0012] According to a preferred embodiment, the pharmaceutically acceptable carrier or excipient is selected from one or more of DMSO, PEG-300, Tween-80, physiological saline, vegetable oil, mannitol, starch, lactose, magnesium stearate, and hydroxypropyl methylcellulose.

[0013] According to a preferred embodiment, the dosage form of the drug is tablets, capsules, granules, pills, powders, granules, ointments, suspensions, oral liquids, or injections.

[0014] According to a preferred embodiment, the injectable is an intradermal injection, a subcutaneous injection, or an intravenous injection.

[0015] According to a preferred embodiment, the dosage of the drug is 10-50 mg / kg / day, preferably 5 mg / kg, 15 mg / kg, 25 mg / kg, 35 mg / kg or 50 mg / kg.

[0016] According to a preferred embodiment, the psoriasis includes psoriasis vulgaris, erythrodermic psoriasis, psoriatic arthritis, or pustular psoriasis.

[0017] This invention also provides an intradermal injection solution for treating psoriasis, comprising a small molecule compound Z57346765 or a pharmaceutically acceptable salt thereof, a solvent, and pharmaceutically acceptable excipients; preferably, the solvent comprises DMSO, PEG-300, and Tween-80, with a preferred volume ratio of 1:5:4; the concentration of the small molecule compound Z57346765 or a pharmaceutically acceptable salt thereof in the injection solution is 5-25 mg / mL; wherein, the structural formula of the small molecule compound Z57346765 is: .

[0018] This invention also provides a topical pharmaceutical composition for treating psoriasis, comprising an effective dose of a small molecule compound Z57346765 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable complex carrier, wherein the structural formula of the small molecule compound Z57346765 is: .

[0019] Z57346765 is a small molecule compound with good transdermal absorption potential. Its small molecular weight and moderate lipid solubility allow it to penetrate the skin's stratum corneum barrier and reach the lesion site to exert its effect. In this example, the injectable solution is administered directly to the lesion site via intradermal injection. While the topical preparation is applied to the skin surface, its small molecule characteristics allow the drug to passively diffuse into the epidermis and dermis, essentially consistent with the "local targeting" effect of injection. The only difference is the route of administration; it does not affect the binding and action of the drug to the target PGK1. The core lesions of psoriasis are concentrated in the local skin (epidermal hyperplasia and dermal inflammatory infiltration). The lesions are superficial, and the target can be reached without systemic administration. In this example, the injectable solution delivers the drug directly to the lesion site via intradermal injection, while the topical preparation can directly cover the lesion surface. After transdermal penetration, the drug can form an effective concentration at the lesion site, consistent with the "local drug accumulation" effect of injection. Current mainstream treatments for psoriasis include topical medications (such as corticosteroid creams and vitamin D analogue ointments), which have demonstrated the feasibility of "local drug delivery targeting local skin lesions." The core mechanism of action of this invention is that Z57346765 specifically inhibits the expression or activity of PGK1, thereby inhibiting excessive proliferation of epidermal keratinocytes and reducing inflammatory responses. The key to this mechanism lies in the "binding of the drug to PGK1," regardless of the drug delivery method (injection / topical application). Whether injected directly into the lesion or applied transdermally, as long as the drug reaches the lesion site and maintains an effective concentration, it can exert its PGK1 inhibitory effect. Examples have confirmed that Z57346765 effectively treats psoriasis by inhibiting PGK1, while topical administration only changes the route of drug entry into the body, without altering the drug's target and mechanism of action. Compared with existing technologies, this invention has the following beneficial effects: The discovery of the small molecule compound Z57346765 with psoriasis therapeutic activity expands the application field of this compound and provides a new drug option for the treatment of psoriasis.

[0020] Targeting PGK1, a novel therapeutic target for psoriasis, breaks through the traditional approach of existing drugs that mainly target inflammatory factors. By regulating metabolic reprogramming, it intervenes in psoriasis at the pathogenesis level, resulting in stronger targeting and more lasting efficacy.

[0021] Experimental results show that Z57346765 can significantly improve symptoms of erythema, scaling and epidermal thickening in psoriatic mice, reduce PASI scores, and has a simple administration method and clear local effects, showing good prospects for clinical application. Attached Figure Description

[0022] Figure 1Image of skin lesions on the back of mice; Part A is the imiquimod model group (IMQ group), and Part B is the Z57346765 treatment group (IMQ+Z57346765 group), showing that the erythema and scaling symptoms of the skin lesions were significantly improved in the treatment group.

[0023] Figure 2 : Trend chart of PASI score for skin lesions on the back of mice; the horizontal axis represents the number of days of administration, and the vertical axis represents the PASI score. It shows that the PASI score of the Z57346765 treatment group was significantly lower than that of the IMQ group, and it showed a decreasing trend with the extension of administration time.

[0024] Figure 3 : Comparison of HE staining and epidermal thickness of mouse skin tissue; where A and B are HE stained sections (scale bar is 100μm), and C is a bar chart of epidermal thickness. (P<0.001) indicates that the epidermal thickness in the treatment group was significantly lower than that in the IMQ group, and the difference was statistically significant (Part C). Detailed Implementation

[0025] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0026] Unless otherwise specified, the methods described in the following examples are conventional methods; unless otherwise specified, the reagents and materials used are conventional reagents and materials in the art and can be obtained commercially.

[0027] I. Experimental Materials Experimental animals: Twelve female C57BL / 6 mice of equal weight, aged 6-8 weeks, were purchased from Beijing Huafukang Biotechnology Co., Ltd., and housed in an SPF-grade animal room at a temperature of 22-25℃ and a humidity of 50-60%, with free access to food and water.

[0028] Experimental drugs and reagents: Small molecule compound Z57346765 (CAS#1016340-64-9, purity: 95%, purchased from MCE, Shanghai, China); Imiquimod cream (National Drug Approval Number H20030129, purchased from Sichuan Mingxin Pharmaceutical Co., Ltd.); DMSO, PEG-300, Tween-80 (analytical grade, all purchased from MCE, Shanghai, China); physiological saline (0.9% NaCl solution); 4% paraformaldehyde fixative, hematoxylin-eosin (HE) staining kit (purchased from Beyotime, catalog number C0105S).

[0029] Experimental instruments: electronic balance, small animal hair clipper, syringe (1mL), pathological slicer, microscope, image analysis software (ImageJ).

[0030] II. Experimental Methods Construction and grouping of a mouse model of psoriasis Twelve C57BL / 6 mice were randomly divided into an IMQ modeling group (control group) and a Z57346765 treatment group, with six mice in each group. After three days of acclimatization, the skin on the back of the mice was shaved off using a small animal shaver, covering an area of ​​2cm × 3cm, ensuring that the skin remained intact and undamaged.

[0031] Modeling method: 62.5 mg of imiquimod cream was applied to the bare skin on the back of both groups of mice daily for 5 consecutive days to induce the formation of psoriasis-like lesions.

[0032] Administration method: On days 1, 2, 4 and 5 after modeling, mice in the treatment group were injected intradermally with Z57346765 solution (dose of 25 mg / kg / day) at the skin lesions on their backs. The volume ratio of the solvent was 10% DMSO: 40% PEG300: 5% Tween-80: 45% saline. The control group was injected intradermally with the same volume of solvent (a saline solution prepared by DMSO, PEG-300 and Tween-80 in the same proportion).

[0033] Skin lesion assessment (PASI score) Starting from day 1 of modeling, the skin lesions on the backs of mice were observed and recorded daily. PASI scores were calculated according to the following criteria, with scoring indicators including erythema, infiltration, and scaling. Each indicator was scored from 0 to 4 points, and the total score was the sum of the scores for the three indicators. The scoring criteria are shown in the table below:

[0034] Sample collection and processing Twenty-four hours after administration on day 5 of modeling, mice were euthanized by cervical dislocation, and a skin lesion tissue sample of approximately 1cm × 1cm × 0.2cm was obtained from the back using a scalpel, ensuring the integrity of the dermis.

[0035] The tissue samples were divided into two parts: one part was immediately placed in a fixative containing 4% paraformaldehyde (the volume of the fixative was more than 10 times the volume of the tissue) and fixed at room temperature for 12 hours for HE staining; the other part was flash-frozen in liquid nitrogen and then transferred to a -80℃ freezer for storage for subsequent PGK1 expression level detection.

[0036] HE staining and epidermal thickness analysis (1) Dehydration and transparency: The fixed tissue was soaked in 70% ethanol for 2 hours, 80% ethanol for 1 hour, 95% ethanol for 1 hour (replaced twice), and anhydrous ethanol for 30 minutes (replaced twice) to completely remove the tissue water; then the tissue was immersed in xylene for 15 minutes each time (replaced twice) to make the tissue transparent.

[0037] (2) Paraffin infiltration and embedding: After the tissue has been transparent, it is placed in molten paraffin at 56-58℃ and soaked for 1 hour (replaced three times) to allow the paraffin to fully penetrate the tissue; after the tissue has been infiltrated, it is placed in an embedding mold, the position is adjusted and then molten paraffin is poured in, and the tissue is placed in cold water to allow the paraffin to solidify into a block quickly.

[0038] (3) Slicing and spreading: Use a microtome to cut the paraffin-embedded block into continuous slices with a thickness of 5μm. Place the slices in warm water at 40-45℃ to spread them naturally. Use a glass slide to lift them out and place them on a 60℃ baking machine to bake for 1 hour to make the slices adhere firmly.

[0039] (4) HE staining: Soak the baked sections in xylene for 5 minutes (replace twice) to dewax; hydrate in sequence by passing through anhydrous ethanol for 5 minutes, 95% ethanol for 3 minutes, 80% ethanol for 3 minutes, and 70% ethanol for 3 minutes, and rinse with distilled water for 2 minutes; stain with hematoxylin for 5 minutes, rinse with tap water for 1 minute, differentiate with 1% hydrochloric acid alcohol for a few seconds, and then rinse with tap water to return to blue for 5 minutes; stain with eosin for 3 minutes, and rinse with tap water; dehydrate in sequence by passing through 70% ethanol, 80% ethanol, and 95% ethanol for 2 minutes each, and anhydrous ethanol for 5 minutes; soak in xylene for 5 minutes (replace twice) to clear; mount with neutral resin to avoid the formation of air bubbles.

[0040] (5) Microscopic examination and analysis: Observe the stained sections under a microscope, select 5 representative fields of view, measure the epidermal thickness using ImageJ software, and calculate the average epidermal thickness of each group.

[0041] Statistical analysis All experimental data were statistically analyzed using SPSS 23.0 software. Quantitative data were expressed as mean ± standard deviation. The t-test was used for comparisons between groups, and P < 0.05 was considered statistically significant.

[0042] III. Experimental Results Improvement in the appearance of skin lesions like Figure 1 As shown, the IMQ model group mice showed obvious erythema, thickening and a large amount of scales on the back of their skin, which is consistent with the characteristics of psoriasis-like skin lesions; the mice in the Z57346765 treatment group showed lighter erythema color, reduced lesion elevation, significantly reduced scales, and significant improvement in the appearance of skin lesions.

[0043] PASI score results like Figure 2 As shown, during the modeling period, the PASI score of the IMQ modeling group continued to increase; the PASI score of the Z57346765 treatment group was significantly lower than that of the control group, and the difference between the groups was statistically significant (P<0.001).

[0044] Epidermal thickness analysis results like Figure 3 As shown, HE staining results indicated that the epidermis in the IMQ model group was significantly thickened, with hyperkeratosis and parakeratosis; the epidermal thickness in the Z57346765 treatment group (PGK1 inhibitor group) was significantly reduced, and the difference was statistically significant (P<0.05).

[0045] Experimental data from the IMQ model group and the PGK1 inhibitor group

[0046] IV. Experimental Conclusions This embodiment validates, using an imiquimod-induced psoriasis mouse model, that intradermal injection of the small molecule compound Z57346765 at a dose of 25 mg / kg / day significantly improves skin lesions, reduces PASI scores and epidermal thickness in psoriasis mice, demonstrating a clear therapeutic effect on psoriasis. These results indicate that Z57346765 can be used as an active ingredient in the preparation of psoriasis treatment drugs, providing a new option for clinical psoriasis treatment.

Claims

1. The application of small molecule compound Z57346765 and its pharmaceutically acceptable salt in the preparation of drugs for treating psoriasis, wherein, The structural formula of the small molecule compound Z57346765 is as follows: 。 2. The application according to claim 1, characterized in that, The Z57346765 and its pharmaceutically acceptable salts, or the drug, exert their therapeutic effect on psoriasis by inhibiting the expression or activity of PGK1.

3. The application according to claim 1, characterized in that, The drug comprises an effective dose of the small molecule compound Z57346765 and a pharmaceutically acceptable carrier or excipient.

4. The application according to claim 3, characterized in that, The pharmaceutically acceptable carrier or excipient is selected from one or more of DMSO, PEG-300, Tween-80, physiological saline, vegetable oil, mannitol, starch, lactose, magnesium stearate, and hydroxypropyl methylcellulose.

5. The application according to claim 1, characterized in that, The dosage form of the drug is tablets, capsules, granules, pills, powders, granules, ointments, suspensions, oral liquids, or injections.

6. The application according to claim 5, characterized in that, The injectable is an intradermal injection, subcutaneous injection, or intravenous injection.

7. The application according to claim 1, characterized in that, The dosage of the drug is 5-50 mg / kg / day, preferably 5 mg / kg, 15 mg / kg, 25 mg / kg, 35 mg / kg or 50 mg / kg.

8. The application according to claim 1, characterized in that, The psoriasis mentioned includes plaque psoriasis, erythrodermic psoriasis, psoriatic arthritis, or pustular psoriasis.

9. An intradermal injection solution for treating psoriasis, characterized in that, This includes small molecule compound Z57346765 or its pharmaceutically acceptable salts, solvents, and pharmaceutically acceptable excipients; Preferably, the solvent includes DMSO, PEG-300 and Tween-80, and the volume ratio of the three is preferably 1:5:4; the concentration of the small molecule compound Z57346765 or its pharmaceutically acceptable salt in the injection solution is 5-25 mg / mL; The structural formula of the small molecule compound Z57346765 is as follows: 。 10. A topical pharmaceutical composition for treating psoriasis, characterized in that, The invention comprises an effective dose of a small molecule compound Z57346765 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable complex carrier, wherein the structural formula of the small molecule compound Z57346765 is: 。

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