Use of a phgdh inhibitor, nct-503, in the preparation of a medicament for preventing and treating parkinson's disease

Abstract

The application belongs to the technical field of medicine and the technical field of brain degenerative diseases, and particularly relates to application of a PHGDH inhibitor NCT-503 in preparation of a medicine for preventing and treating Parkinson's disease, and verifies that specific inhibition of PHGDH can inhibit nerve inflammation in the brain substantia nigra region in vivo, reduce death of dopaminergic neurons, and thus relieve occurrence and development of Parkinson's disease; acute Parkinson's disease model mice are constructed by intraperitoneal injection of MPTP, and it is found that intraperitoneal injection treatment of NCT-503 can effectively reduce MPTP-induced nerve inflammation and reduce death of dopaminergic neurons. In conclusion, it is found for the first time that use of NCT-503 to inhibit PHGDH activity can significantly inhibit MPTP-induced nerve inflammation and reduce death of dopaminergic neurons, which provides an important theoretical basis for clinical application of NCT-503, and also provides a new idea for medicine research and development and screening for treating Parkinson's disease.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine and the technical field of brain degenerative diseases, and particularly relates to application of a PHGDH inhibitor NCT-503 in preparation of a medicine for preventing and treating Parkinson's disease. BACKGROUND

[0002] Parkinson's disease, often referred to as "paralysis agitans", is a neurodegenerative disease. The main cause of this disease is the degeneration and death of dopaminergic neurons in the substantia nigra, which may be related to multiple factors such as genetics, environmental factors and aging of the nervous system. Parkinson's disease symptoms vary from person to person, and the most prominent symptoms are static tremor, bradykinesia and muscle rigidity. Patients in the middle and late stages have posture balance disorders, and some non-motor symptoms may occur before and after the onset, including constipation, olfactory dysfunction, sleep disorders, autonomic nervous dysfunction and mental and cognitive disorders. Studies have found that neuroinflammation plays an important role in the progression of Parkinson's disease, so intervention and regulation of neuroinflammation may provide a new way for the treatment of Parkinson's disease.

[0003] 3-phosphoglycerate dehydrogenase (PHGDH) is the first rate-limiting enzyme in the serine synthesis pathway, which is mainly responsible for converting the glycolysis intermediate 3-phosphoglycerate (3-PG) into 3-phosphohydroxypyruvate (3-PHP), and ultimately entering the one-carbon cycle to maintain the corresponding material homeostasis. The applicant team found that PHGDH-mediated astrocyte inflammatory response plays an important regulatory role in Parkinson's disease, but there is currently no research report on inhibiting the progression of Parkinson's disease by inhibiting PHGDH activity. SUMMARY

[0004] The purpose of the present application is to provide a PHGDH inhibitor NCT-503 for use in the preparation of a medicine for preventing and treating Parkinson's disease, which can significantly inhibit 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neuroinflammation and reduce MPTP-induced death of dopaminergic neurons, thereby providing an important theoretical basis for the clinical application of NCT-503.

[0005] The purpose of the present application is achieved by the following technical solutions:

[0006] The present application provides a PHGDH inhibitor NCT-503 for use in the preparation of a medicine for preventing and treating Parkinson's disease.

[0007] Further, the PHGDH inhibitor NCT-503 prevents and treats Parkinson's disease by inhibiting PHGDH activity.

[0008] Further, the inhibition of PHGDH activity can improve the survival of dopaminergic neurons in mice after MPTP, reduce the MPTP-induced reactive astrocytes, and reduce the expression of inflammatory factors in the tissue of the substantia nigra region of the brain.

[0009] Further, the inflammatory factors are TNFα, IL-1β and IL-6.

[0010] Further, the dosage form of the drug includes tablets, capsules, oral agents and injections.

[0011] Further, the tablets include coated tablets.

[0012] Further, the oral agents include oral liquid agents, oral granules and oral powders.

[0013] Further, the injections include lyophilized powders and emulsions for injection. Advantages

[0014] The present application verifies that specific inhibition of PHGDH can inhibit neuroinflammation in the substantia nigra region of the brain in vivo, thereby regulating the occurrence and development of Parkinson's disease; the present application constructs an acute Parkinson's disease model mouse by intraperitoneal injection of MPTP in C57BL / 6 mice, and the results show that intraperitoneal injection of NCT-503 can effectively reduce MPTP-induced neuroinflammation and reduce the death of dopaminergic neurons. In summary, the present application first finds that the use of NCT-503 to inhibit PHGDH activity can significantly inhibit MPTP-induced neuroinflammation and reduce the death of dopaminergic neurons, thereby providing an important theoretical basis for the clinical application of NCT-503 and providing a new idea for the research and screening of drugs for treating Parkinson's disease. BRIEF DESCRIPTION OF DRAWINGS

[0015] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed in the embodiments. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings according to these drawings without creative labor.

[0016] Figure 1 is the specific experimental flowchart in Example 1 of the present application;

[0017] Figure 2 is the TH expression in normal mice treated with NCT-503 in Example 2 of the present application; + The graph of the influence of neuron survival, wherein the left graph is the fluorescence staining result graph, and the right graph is the columnar graph of the influence results of different groups;

[0018] Figure 3Figure 4 is a graph showing the effect of NCT-503 on TH expression in the brain of a mouse with Parkinson's disease (acute MPTP model) according to Example 3 of the present application. + Figure 5 is a neuron survival impact graph, in which the left graph is a fluorescence staining result graph, and the right graph is a column graph of the impact results of different groups;

[0019] Figure 4 Figure 6 is a graph showing the effect of NCT-503 on astrocyte activation in the brain of a mouse with Parkinson's disease (acute MPTP model) according to Example 4 of the present application, in which the left graph is a fluorescence staining result graph, and the right graph is a column graph of the impact results of different groups;

[0020] Figure 5 Figure 7 is a graph showing the effect of NCT-503 on the expression of inflammatory factors in the brain of a mouse with Parkinson's disease (acute MPTP model) according to Example 5 of the present application, in which the inflammatory factors are TNFα, IL-1β, and IL-6 in turn. DETAILED DESCRIPTION

[0021] The following detailed description of various example embodiments of the present application is not to be taken in a limiting sense but is made with reference to the drawings in which like numerals represent like elements and in which:

[0022] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the present application. In addition, any numerical range recited herein is intended to include all sub-ranges of the same numerical precision subsumed therein. In addition, all listed numerical values are "approximate", meaning that the term "comprises" to be read equivalently to "from" or "from about". In this document, the terms "comprises" and variations thereof do not have a limiting meaning, such that a composition or method that comprises a list of elements is not necessarily limited to only those elements but can include other elements not expressly listed or inherent to such composition or method. In addition, unless expressly stated to the contrary, the term "or" as used herein refers to an inclusive "or" and not to an exclusive "or". That is, unless specifically stated to the contrary, "or" means allowing for "either or" or "both".

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, preferred methods and materials are described. All publications mentioned herein are incorporated by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present application is not entitled to antedate such publication by virtue of prior application.

[0024] Various modifications and changes can be made to the specific embodiments of the present application described herein without departing from the scope or spirit of the application. Other embodiments of the application will be apparent to those of ordinary skill in the art from the description and examples presented herein. The description and examples are illustrative of the application and are not intended to limit the scope of the application in any way.

[0025] As used herein, the terms “comprises,” “comprising,” “includes,” “including,” “has,” “having,” and the like are open-ended terms that are intended to be synonymous with each other, and are used to include, but not limited to, the stated item.

[0026] The experimental methods used in the following examples are the commonly used experimental methods in the art, unless otherwise specified.

[0027] The experimental materials used in the following examples are the commonly used experimental materials in the art, unless otherwise specified.

[0028] In the following examples, the C57BL / 6 mice used were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., license number: SYXK (Lu) 20220006. The mice were raised in the Experimental Animal Platform of the Biomedical Center of Qingdao University, and the experiment was carried out after 7 days of adaptive feeding; the NCT-503 used was purchased from Tao Shu Biotechnology (Shanghai) Co., Ltd.

[0029] Example 1

[0030] In this example, C57BL / 6 mice were injected intraperitoneally with MPTP (20 mg / kg) at 0 hours, 2 hours, 4 hours, and 6 hours to induce an acute Parkinson's disease model, and normal saline was used as a control. Then, NCT-503 or its solvent (solvent components include 5% ethanol + 35% PEG300 + 60% HBC) was injected at 24 hours, 48 hours, and 72 hours after MPTP injection, and 7 days later, immunofluorescence staining of tissues or cytokine detection was performed. The specific experimental flowchart is shown in Figure 1 .

[0031] Example 2 NCT-503 on TH + Neuron survival impact

[0032] In this example, NCT-503 and vehicle solvent (Vehicle) were injected intraperitoneally in C57BL / 6 mice under physiological conditions to verify the effect of NCT-503 on TH+ neuron survival in the substantia nigra region under normal conditions in vivo. Then, mouse brain frozen sections were taken and stained with dopamine neuron marker protein antibody (anti-TH) for fluorescence staining to evaluate the effect of NCT-503 on dopamine neurons under physiological conditions.

[0033] From Figure 2 As can be seen from the results, the administration of NCT-503 or its solvent under normal conditions does not affect the number of TH-positive neurons in the substantia nigra region of mice. Injection of saline (Saline), NCT-503, and solvent (Vehicle) under physiological conditions in mice has no significant effect on the survival rate of dopamine neurons.

[0034] Example 3 Effect of NCT-503 on TH survival in Parkinson's disease mice (acute MPTP model) + Effect on neuron survival

[0035] In this example, the effect of NCT-503 on TH survival in Parkinson's disease mice (acute MPTP model) was investigated. + The specific experimental procedure for the effect of NCT-503 on neuron survival was as follows:

[0036] (1) The mice were induced to an acute MPTP model (MPTP was injected intraperitoneally, 20 mg / kg, injected every 2 hours for 4 times), and randomly divided into an experimental group (NCT-503) and a solvent control group (Vehicle), which were treated with NCT-503 and Vehicle, respectively (intraperitoneal injection at 24, 48 and 72 hours after MPTP induction, with a dose of 20 mg / kg each time), and 4 C57BL / 6 mice without any treatment were taken as normal blank controls Control.

[0037] (2) After 7 days of MPTP injection, brain frozen sections were taken and fluorescently stained with dopamine neuron marker protein antibody (anti-TH) to evaluate the effect of NCT-503 treatment on the survival rate of dopamine neurons during Parkinson's disease.

[0038] As can be seen from Figure 3 The results showed that NCT-503 treatment could significantly improve the survival of dopamine neurons in mice after MPTP.

[0039] Example 4 Effect of NCT-503 on astrocyte activation in Parkinson's disease mice (acute MPTP model)

[0040] In this example, the experimental procedure for investigating the effect of NCT-503 on astrocyte activation in Parkinson's disease mice (acute MPTP model) was as follows:

[0041] (1) The experimental conditions were as shown in step (1) of Example 3;

[0042] (2) After 7 days of MPTP injection, brain frozen sections were taken and fluorescently stained with astrocyte marker GFAP to evaluate the effect of NCT-503 treatment on astrocyte activation during Parkinson's disease by detecting the fluorescence intensity of GFAP, as shown in Figure 4 .

[0043] As can be seen from Figure 4 , NCT-503 treatment could significantly reduce the astrocyte reactivity induced by MPTP.

[0044] Example 5 Effect of NCT-503 on expression of inflammatory factors in the substantia nigra region of the brain of a mouse with Parkinson's disease (acute MPTP model)

[0045] (1) The experimental conditions were as shown in step (1) of Example 3.

[0046] (2) On the seventh day after induction of the MPTP model, the mice were killed and the brains were removed for RNA extraction. qRT-PCR was performed on the prepared brain tissue RNA samples to detect the expression of inflammatory factors (TNFα, IL-1β, IL-6) in the brain tissue of the experimental and control groups.

[0047] The results are shown in Table 1. Figure 5 As shown in Table 1, NCT-503 treatment significantly reduced the expression of inflammatory factors in the substantia nigra region of the brain of the mouse.

[0048] In summary, the PHGDH-specific inhibitor NCT-503 used in the present application can significantly improve the survival rate of dopaminergic neurons in the substantia nigra region of a mouse with MPTP-induced Parkinson's disease, and can significantly inhibit the neuroinflammation mediated by astrocytes during the development of Parkinson's disease. This finding provides an important theoretical basis for the clinical application of NCT-503 and also provides a new approach for the development and screening of drugs for the treatment of Parkinson's disease.

[0049] The above examples only express several embodiments of the present application, and the description is relatively specific and detailed, but it should not be understood as limiting the scope of the patent. It should be noted that, for those skilled in the art, without departing from the concept of the present application, several modifications and improvements can be made, which are all within the scope of the present application. Therefore, the scope of protection of the present application should be subject to the appended claims.

Claims

1. Use of a PHGDH inhibitor NCT-503 in the preparation of a drug for preventing and treating Parkinson's disease; The dosage forms of the drug include tablets, capsules, oral liquids, oral granules, oral powders and injections; The tablets include coated tablets; The injection includes freeze-dried powder injection and injection emulsion.

2. The use according to claim 1, characterized in that The PHGDH inhibitor NCT-503 prevents and treats Parkinson's disease by inhibiting PHGDH activity.

3. The use according to claim 2, characterized in that The inhibition of PHGDH activity can improve the survival of dopaminergic neurons in mice after MPTP injection, reduce the MPTP-induced astrocyte reactivity, and reduce the expression of inflammatory factors in the substantia nigra region of the brain.

4. The use according to claim 3, characterized in that The inflammatory factors are TNFα, IL-1β and IL-6.

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