Application of PHGDH inhibitor NCT-503 in preparation of medicine for preventing and treating Parkinson's disease

Abstract

The invention belongs to the technical field of medicines 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, which verifies that specific inhibition of PHGDH can inhibit neuroinflammation in a brain nigra area in vivo, relieve death of dopaminergic neurons and prevent and treat Parkinson's disease. Therefore, occurrence and development of Parkinson's disease are relieved; an acute Parkinson's disease model mouse is constructed through intraperitoneal injection of MPTP, and it is found that NCT-503 intraperitoneal injection treatment can effectively relieve MPTP-induced neuroinflammation and relieve death of dopaminergic neurons. In conclusion, it is found for the first time that the NCT-503 is used for inhibiting the activity of the PHGDH, MPTP-induced neuroinflammation can be remarkably inhibited, and death of dopaminergic neurons is relieved, an important theoretical basis is provided for clinical application of the NCT-503, and a new thought is provided for research, development and screening of drugs for treating the Parkinson's disease.

Description

Technical Field

[0001] The present invention belongs to the field of medicine technology and brain degenerative disease technology, and specifically relates to the use of a PHGDH inhibitor NCT-503 in the preparation of a drug for preventing and treating Parkinson's disease. Background Art

[0002] Parkinson's disease, also often referred to as "tremor palsy", is a neurodegenerative disease. The main cause of this disease is due to 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. The symptoms of Parkinson's disease vary from person to person. The most significant symptoms are resting tremor, bradykinesia and muscle rigidity. Patients in the middle and late stages have postural balance disorders. There are also some non-motor symptoms before and after the onset of the disease, including constipation, olfactory disorders, sleep disorders, autonomic dysfunction, and mental and cognitive disorders. Studies have found that neuroinflammation plays an important role in the progression of Parkinson's disease. Therefore, intervention and regulation of neuroinflammation may provide a new way to treat Parkinson's disease.

[0003] 3-Phosphoglycerate dehydrogenase (PHGDH) is the first rate-limiting enzyme in the serine biosynthesis pathway. It is mainly responsible for converting the glycolysis intermediate 3-phosphoglycerate (3-PG) into 3-phosphohydroxypyruvate (3-PHP), which eventually enters the one-carbon cycle, thereby maintaining the corresponding material homeostasis. The applicant team found that the PHGDH-mediated astrocyte inflammatory response plays an important regulatory role in Parkinson's disease, but there are no research reports on inhibiting the progression of Parkinson's disease by inhibiting PHGDH activity at home and abroad. Summary of the invention

[0004] The purpose of the present invention is to provide a PHGDH inhibitor NCT-503 for use in the preparation of drugs 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 dopaminergic neuron death, providing an important theoretical basis for the clinical application of NCT-503.

[0005] The objective of the present invention is achieved through the following technical solutions: The present invention provides use of a PHGDH inhibitor NCT-503 in preparing a drug for preventing and treating Parkinson's disease.

[0006] Furthermore, the PHGDH inhibitor NCT-503 prevents and treats Parkinson's disease by inhibiting PHGDH activity.

[0007] Furthermore, the inhibition of PHGDH activity can improve the survival of dopaminergic neurons in mice after MPTP, reduce the reactivity of astrocytes induced by MPTP, and reduce the expression of inflammatory factors in the substantia nigra region of the brain.

[0008] Furthermore, the inflammatory factors are TNFα, IL-1β and IL-6.

[0009] Furthermore, the dosage forms of the drug include tablets, capsules, oral preparations and injections.

[0010] Furthermore, the tablet comprises a coated tablet.

[0011] Furthermore, the oral preparation includes oral liquid, oral granules and oral powder.

[0012] Furthermore, the injection includes freeze-dried powder injection and injection emulsion. Beneficial Effects

[0013] The present invention 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 invention constructs acute Parkinson's disease model mice 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 invention first discovered that the use of NCT-503 to inhibit PHGDH activity can significantly inhibit MPTP-induced neuroinflammation and reduce the death of dopaminergic neurons, which provides an important theoretical basis for the clinical application of NCT-503, and also provides new ideas for the development and screening of drugs for the treatment of Parkinson's disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0015] Figure 1 This is a specific experimental flow chart in Example 1 of the present invention; Figure 2 The effect of NCT-503 on TH in normal mice in Example 2 of the present invention + Neuron survival impact diagram, where the left picture is the fluorescence staining result diagram, and the right picture is the bar graph of the impact results of different groups; Figure 3 The effect of NCT-503 on TH in Parkinson's disease mice (acute MPTP model) in Example 3 of the present invention+ Neuron survival impact diagram, where the left picture is the fluorescence staining result diagram, and the right picture is the bar graph of the impact results of different groups; Figure 4 This is a diagram showing the effect of NCT-503 on astrocyte activation in Parkinson's disease mice (acute MPTP model) in Example 4 of the present invention, wherein the left figure is a fluorescence staining result diagram, and the right figure is a bar graph of the effect results of different groups; Figure 5 This is a graph showing the effect of NCT-503 in Example 5 of the present invention on the expression of inflammatory factors in the substantia nigra region of Parkinson's disease mice (acute MPTP model), wherein the inflammatory factors are TNFα, IL-1β, and IL-6, respectively. DETAILED DESCRIPTION

[0016] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0017] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0018] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0019] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0020] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0021] The test methods used in the following implementations, unless otherwise specified, are all commonly used experimental methods in this field; Unless otherwise specified, the experimental materials used in the following examples are all commonly used experimental materials in this field.

[0022] The C57BL / 6 mice used in the following examples 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 experiments were carried out after 7 days of adaptive breeding; the NCT-503 used was purchased from Taoshu Biotechnology (Shanghai) Co., Ltd.

[0023] Example 1 In this example, MPTP (20 mg / kg) was injected into the abdominal cavity of C57BL / 6 mice at 0 hours, 2 hours, 4 hours and 6 hours to induce an acute Parkinson's disease model and normal saline as a control. Then, NCT-503 or its solvent (the solvent composition includes 5% ethanol (ethanol) + 35% PEG300 (polyethylene glycol) + 60% HBC (cyclodextrin)) was injected as a control 24 hours, 48 ​​hours and 72 hours after the injection of MPTP. Immunofluorescence staining or cytokine detection of tissues was performed 7 days later. The specific experimental flow chart is shown in Figure 1 .

[0024] Example 2 Effect of NCT-503 on TH in Normal Mice + Effects on neuronal survival In this example, NCT-503 and a vehicle solvent (Vehicle) were intraperitoneally injected into C57BL / 6 mice under physiological conditions to verify the effect of NCT-503 on the survival of TH+ neurons in the substantia nigra region under normal in vivo conditions. Frozen sections of the mouse brain were then taken and fluorescently stained with a dopaminergic neuron marker protein antibody (anti-TH) to evaluate the effect of NCT-503 on dopamine neurons under physiological conditions.

[0025] from Figure 2 The results show that under normal circumstances, the administration of NCT-503 or its solvent does not affect the number of TH-positive neurons in the substantia nigra region of mice. Under physiological conditions, the injection of saline (Saline), NCT-503 and solvent (Vehicle) into mice has no significant effect on the survival rate of dopamine neurons.

[0026] Example 3 Effect of NCT-503 on TH in Parkinson's disease mice (acute MPTP model) + Effects on neuronal survival This example investigates the effect of NCT-503 on TH in Parkinson's disease mice (acute MPTP model).+ The specific experimental procedure for the effect of neuronal survival is as follows: (1) Induced acute MPTP model in mice (MPTP was intraperitoneally injected at an interval of 2 hours for 4 times) and randomly divided into an experimental group (NCT-503) and a solvent control group (Vehicle). NCT-503 and Vehicle were used for treatment (intraperitoneal injection at 24, 48, and 72 hours after MPTP induction, with each injection at a dose of 20 mg / kg). Four C57BL / 6 mice that were not treated were used as normal blank controls. (2) Seven days after MPTP injection, brain frozen sections were obtained and fluorescently stained with dopaminergic neuron marker protein antibody (anti-TH) to evaluate the effect of NCT-503 treatment on the survival rate of dopaminergic neurons in Parkinson's disease.

[0027] from Figure 3 The results showed that NCT-503 treatment could significantly improve the survival of dopaminergic neurons after MPTP in mice.

[0028] Example 4 Effect of NCT-503 on astrocyte activation in Parkinson's disease mice (acute MPTP model) The experimental process for investigating the effect of NCT-503 on astrocyte activation in Parkinson's disease (acute MPTP model) in mice in this example is as follows: (1) The experimental conditions are as shown in step (1) in Example 3; (2) Seven days after MPTP injection, brain frozen sections were obtained and fluorescently stained with the astrocyte marker GFAP. The effect of NCT-503 treatment on astrocyte activation in the process of Parkinson's disease was evaluated by detecting the GFAP fluorescence intensity. The results are as follows: Figure 4 shown.

[0029] from Figure 4 It can be seen that NCT-503 treatment can significantly reduce MPTP-induced astrocyte reactivity.

[0030] Example 5 Effect of NCT-503 on the expression of inflammatory factors in the substantia nigra of Parkinson's disease mice (acute MPTP model) (1) The experimental conditions are as shown in step (1) in Example 3; (2) On the seventh day after MPTP model induction, mice were killed and their brains were taken for brain tissue RNA extraction. The prepared brain tissue RNA samples were subjected to qRT-PCR experiments to detect the expression of inflammatory factors (TNFα, IL-1β, IL-6) in the brain tissues of the experimental and control groups.

[0031] The results are as follows Figure 5 As shown, NCT-503 treatment significantly reduced the expression of inflammatory factors in the substantia nigra region of the mouse brain.

[0032] In summary, the PHGDH-specific inhibitor NCT-503 used in the present invention can significantly improve the survival rate of dopaminergic neurons in the substantia nigra region during MPTP-induced Parkinson's disease in mice, and can significantly inhibit astrocyte-mediated neuroinflammation during Parkinson's disease. This discovery provides an important theoretical basis for the clinical application of NCT-503, and also provides new ideas for the development and screening of drugs for the treatment of Parkinson's disease.

[0033] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. Use of a PHGDH inhibitor NCT-503 in the preparation of drugs for preventing and treating Parkinson's disease.

2. The use according to claim 1, It is 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, It is characterized in that The inhibition of PHGDH activity can improve the survival of dopaminergic neurons in mice after MPTP injection, reduce the reactivity of astrocytes induced by MPTP, and reduce the expression of inflammatory factors in the substantia nigra region tissue.

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

5. The use according to claim 1, It is characterized in that The dosage forms of the drug include tablets, capsules, oral preparations and injections.

6. The use according to claim 5, It is characterized in that The tablets include coated tablets.

7. The use according to claim 5, It is characterized in that The oral preparation includes oral liquid preparation, oral granule preparation and oral powder preparation.

8. The use according to claim 5, It is characterized in that The injection includes freeze-dried powder injection and injection emulsion.

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