Application of PSPH inhibitor in preparation of medicine for preventing, treating or improving nervous system diseases

Abstract

The invention discloses application of a PSPH inhibitor in preparation of a medicine for preventing, treating or improving nervous system diseases, and relates to the technical field of small molecule medicines. The invention provides application of a PSPH (phosphoserine phosphatase) inhibitor in preparation of a medicine for treating nervous system diseases, the activity of the PSPH is inhibited by using a small-molecule chemical inhibitor, the nervous system diseases are treated, especially an excellent anti-epileptic effect is achieved, meanwhile, a small-molecule compound with PSPH inhibition activity is provided, and the application of the small-molecule compound in preparation of the medicine for treating the nervous system diseases in preparation of the medicine for treating the nervous system diseases in preparation of the medicine for treating the nervous system diseases is provided. The blank that no potent PSPH inhibitor exists at present is filled.

Description

Technical Field

[0001] The present invention relates to the technical field of small molecule drugs, and in particular to the use of PSPH inhibitors in preparing drugs for preventing, treating or improving nervous system diseases. Background Art

[0002] The nervous system is the most complex regulatory system in the human body with the most complex structure and function. When the nervous system is diseased, symptoms such as consciousness, cognition, movement disorders and abnormal intracranial pressure often occur, which seriously reduce the patient's quality of life. Common nervous system diseases include cerebrovascular diseases, neurodegenerative diseases, epilepsy, and infectious diseases of the central nervous system.

[0003] Chinese patent CN 112585154A discloses a novel serine derivative compound with improved blood-brain barrier (BBB) ​​permeability and its use, and more specifically, relates to a novel serine derivative compound with improved blood-brain barrier permeability compared with L-serine, and a pharmaceutical composition containing the above compound as an active ingredient for preventing or treating / improving central nervous system diseases. The compound or its pharmaceutically acceptable salt disclosed in the invention shows significantly improved blood-brain barrier permeability compared with L-serine, activates the proliferation of nerve cells, and has a neuroprotective effect by inhibiting the damage of mitochondrial membrane potential caused by oxidative stress and / or neuronal apoptosis caused by endoplasmic reticulum stress. It has excellent effects in preventing, treating and improving central nervous system diseases such as cognitive impairment, intellectual disability, microcephaly, epilepsy, neurodevelopmental disorder, dementia, autism spectrum disorder, Down syndrome, Rett syndrome, fragile X syndrome, Alzheimer's disease, Parkinson's disease, Huntington's disease and amyotrophic lateral sclerosis.

[0004] Excitotoxicity is an important cause of neuronal damage and death in a variety of acute and chronic neuropathies. It is caused by excessive activation of NMDA receptors due to excessive glutamate release or uptake disorders. Studies have shown that D-serine is related to excitotoxicity. For example, the application of D-serine deaminase can significantly reduce neuronal damage and death. The above research results show that D-serine is an endogenous ligand for most NMDA receptors.

[0005] D-serine is found to be unevenly distributed in the brain of animals, with the highest content in the cerebral cortex, hippocampus, and striatum, followed by the diencephalon and midbrain, and the lowest concentration in the pons, medulla oblongata, cerebellum, and spinal cord. There are also reports that D-serine is present in the retina. Studies on mouse brains have shown that the concentration of D-serine is about 1 / 3 of that of L-serine, which is higher than the concentration of many common amino acids. In the human brain, the content of D-serine changes with age, and remains at a high level from 14 weeks of pregnancy until birth, and the concentration is reduced to half in adolescence and mature brains. Compared with glycine, D-serine released by astrocytes is more effective and more selective in binding to the glycine site of NMDA (N-methyl-D-aspartate) receptors as a co-agonist. The binding of D-serine to the glycine site can enhance the stimulatory effect of glutamate on NMDA receptors, leading to postsynaptic membrane depolarization and calcium ion influx.

[0006] Under pathological conditions, excessive activation of NMDA receptors due to excessive D-serine concentrations may be associated with many neurological diseases, such as stroke, epilepsy, chronic pain, Parkinson's disease (PD), Alzheimer's disease (AD), Huntington's disease (DHD), ischemia-reperfusion injury, etc. It has been reported that ischemia-reperfusion injury is alleviated after the administration of glycine site antagonists; the level of D-serine in stroke patients is significantly increased. Although abnormal D-serine concentrations have not been detected in PD and AD patients, the use of glycine site antagonists can improve the behavioral abnormalities of PD patients. In contrast, low NMDA receptor function can lead to schizophrenia. Compared with normal people, the level of D-serine in the serum and prefrontal cortex of such patients is reduced. In epilepsy, the binding of D-serine to the glycine site of the NMDA receptor can produce the opposite result. It has been reported that the activation of NMDA receptors can lead to grand mal seizures; however, other studies have shown that this binding can increase the threshold of grand mal seizures. D-serine has a similar effect on convulsions: on the one hand, NMDA receptor antagonists can inhibit convulsions, and on the other hand, D-serine, as an NMDA receptor co-agonist, can enhance the activity of anticonvulsant drugs. The duality of D-serine's effects may be related to factors such as the different NMDA receptor subtypes it binds to. Its mechanism is not yet fully understood.

[0007] Among neurological diseases, epilepsy is induced by abnormal functional activities of the central nervous system and is one of the most common chronic neurological diseases. Nearly one-third of patients do not respond to anti-epileptic drugs that target ion channels, which is drug-resistant epilepsy. Temporal lobe epilepsy (TLE) is the most common type of drug-resistant epilepsy in adults. After taking enough anti-epileptic drugs correctly, patients still cannot effectively control epileptic seizures, and epilepsy can often only be treated by surgical resection of the hippocampus, the epileptogenic focus. The medical and economic burden that drug-resistant epilepsy brings to society and families is much higher than that of general epilepsy syndromes. Therefore, clarifying the pathogenesis of intractable epilepsy such as temporal lobe epilepsy and developing new and effective therapeutic drugs will provide patients with non-surgical treatment options and fill the treatment gap for intractable epilepsy.

[0008] NMDA is a widely expressed ionotropic glutamate receptor, and its abnormal expression or function plays a crucial role in epileptic discharges. Although small molecule antagonists targeting NMDA (most of which are channel blockers) have shown anti-epileptic effects in clinical practice, they cannot be used as daily medications for TLE patients due to their strong toxic side effects. For example, high doses of ketamine can treat patients in status epilepticus, but it may have effects on the respiratory, cardiovascular and nervous systems. In addition to the glutamate binding site, NMDA also needs to bind to D-serine or glycine at the glycine regulatory site to exert its effect. Studies have shown that D-serine is a co-activator of hippocampal synapses, which affects neuronal electrophysiological activity in a gating regulatory manner. Increased levels of D-serine significantly increase the discharge of hippocampal neurons.

[0009] In the brain, astrocytes synthesize L-serine from glucose through a three-step enzymatic reaction. After being released to the surrounding neurons, D-serine is generated under the action of serine racemase (SR). This mode is called "serine shuttle". Then, D-serine is released to the postsynaptic membrane to act on the NMDAR receptor to further regulate synaptic activity. Studies have shown that inhibitors targeting serine racemase have certain anti-excitatory effects in cell and animal epilepsy models, but existing serine racemase inhibitors do not have the potential for clinical development due to off-target effects and low inhibitory efficacy. On the other hand, blocking the upstream de novo synthesis pathway may be another potential method to reduce D-serine levels and treat temporal lobe epilepsy. However, the molecular cascade reactions and drug targets involved remain to be studied.

[0010] In summary, there is still a need to study small molecule drugs that have effective therapeutic effects on neurological diseases including epilepsy. Summary of the invention

[0011] The purpose of the present invention is to provide the use of PSPH (phosphoserine phosphatase) inhibitors in the preparation of drugs for treating nervous system diseases, using small molecule chemical inhibitors to inhibit PSPH activity, thereby achieving the treatment of nervous system diseases, especially exerting excellent anti-epileptic effects, and at the same time providing small molecule compounds with PSPH inhibitory activity, filling the gap of the current lack of potent PSPH inhibitors.

[0012] In the present invention, compound Z218484536 is the compound 1 described above.

[0013] To achieve the above-mentioned purpose of the invention, the technical solution of the present invention is as follows:

[0014] In a first aspect, the present invention provides use of a PSPH inhibitor in the preparation of a medicament for preventing, treating or ameliorating a nervous system disease.

[0015] The term "PSPH inhibitor" refers to an agent that is able to cross the blood-brain barrier and inhibit the activity of phosphoserine phosphatase target substances.

[0016] Preferably, the PSPH inhibitor prevents, treats or improves nervous system diseases by inhibiting the activity of phosphoserine phosphatase, thereby inhibiting the production and release of L-serine in astrocytes, and thereby inhibiting the upregulation of D-serine.

[0017] Preferably, the nervous system disease is selected from at least one of cognitive impairment, intellectual disability, cerebellar insufficiency, epilepsy, neurodevelopmental disorder, dementia, autism spectrum disorder, Down syndrome, Rett syndrome, fragile X syndrome, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and ischemia-reperfusion injury.

[0018] More preferably, it is at least one of stroke, epilepsy, chronic pain, Parkinson's disease, Alzheimer's disease, Huntington's disease, and ischemia-reperfusion injury.

[0019] More preferably, it is epilepsy.

[0020] More preferably, it is temporal lobe epilepsy.

[0021] Preferably, the PSPH inhibitor is a compound of formula I or a stereoisomer, geometric isomer, tautomer, N-oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof:

[0022]

[0023] In the formula, R1 is selected from H, substituted or unsubstituted C 1-8 Alkyl, C 3-8 Cycloalkyl, aryl, heterocyclic, halogen, C 1-8Alkoxy, the substituted substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6-membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6-membered heterocyclic group containing oxygen and / or nitrogen;

[0024] R2 is -CH=N- or -CH2CH 2- .

[0025] Preferably, R1 is selected from substituted or unsubstituted C 1-5 Alkyl, C 3-5 Cycloalkyl, aryl, 3-6 membered heterocyclic group, halogen, C 1-5 Alkoxy, the substituted substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6-membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6-membered heterocyclic group containing oxygen and / or nitrogen;

[0026] More preferably, R1 is selected from C 1-5 Alkyl, C 3-5 Cycloalkyl, phenyl, halogen, C 1-5 Alkoxy;

[0027] More preferably, R1 is selected from C 1-3 Alkyl, phenyl, halogen, C 1-3 Alkoxy;

[0028] More preferably, R1 is selected from C 1-3 Alkyl, halogen, C 1-3 Alkoxy;

[0029] More preferably, R1 is selected from halogen, C 1-3 Alkoxy;

[0030] More preferably, R1 is selected from F, Cl, Br, C 1-3 Alkoxy;

[0031] More preferably, R1 is selected from F, methoxy; most preferably F

[0032] More preferably, the compound of formula I is selected from the following compounds:

[0033]

[0034] Preferably, the compound of formula I is selected from the following compounds:

[0035]

[0036] Preferably, the drug can be prepared into dosage forms such as pills, capsules, granules, oral liquids, powders, tablets, lozenges, lozenges, injections, etc., and suitable drug carriers in the art can be selected for different dosage forms.

[0037] The pharmaceutical carrier used can be solid, liquid or gas. Examples of solid carriers include lactose, kaolin, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate and stearic acid. Examples of liquid carriers include syrup, peanut oil, olive oil and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0038] When preparing drugs in oral dosage forms, any convenient pharmaceutical medium can be used. For example, water, ethanol, oil, alcohol, flavoring agents, preservatives, colorants, etc. can be used to form oral liquid preparations, such as suspensions, elixirs and solutions; while carriers, such as starch, sugars, microcrystalline cellulose, diluents, granulating agents, emulsifiers, lubricants, binders, disintegrants can be used to form oral solid preparations, such as powders, capsules and tablets. Tablets and capsules are preferred oral dosage units using solid pharmaceutical carriers due to their ease of administration. Tablets can be coated using standard aqueous or non-aqueous techniques.

[0039] Tablets can be prepared by tableting or molding, and one or more auxiliary ingredients or adjuvants can be used optionally. The active ingredient can be tableted in a free-flowing form (such as powder or granules) in a suitable machine, and can be optionally mixed with a binder, lubricant, inert diluent, surfactant or dispersant to prepare a tablet. Molded tablets can be molded in a suitable machine, that is, a powdered compound mixture moistened with an inert liquid diluent. Each tablet preferably contains about 0.05 mg to about 5 g of active ingredient, and each sachet or capsule preferably contains about 0.05 mg to about 5 g of active ingredient. For example, a preparation intended for oral administration to humans may contain about 0.5 mg to about 5 g of active drug, mixed with an appropriate and convenient carrier material, which may account for about 5% to 95% of the total composition. The unit dosage form usually contains about 1 mg to about 2 g of active ingredient, usually 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg or 1000 mg.

[0040] Drugs suitable for parenteral administration may be prepared as aqueous solutions or suspensions of the active compound. Suitable surfactants may be included, such as hydroxypropylcellulose. Dispersions may also be prepared in glycerol, liquid polyethylene glycols, and oil mixtures thereof. In addition, preservatives may be added to prevent the harmful growth of microorganisms.

[0041] The medicament of the present invention can be in a form suitable for topical use, such as an aerosol, cream, ointment, lotion, powder or the like. In addition, the composition can be in a suitable form for transdermal administration devices. For example, a cream or ointment with a desired consistency is prepared by mixing a hydrophilic material and water, and about 5wt% to about 10wt% of the compound.

[0042] The medicine of the present invention can be in a form suitable for rectal administration, wherein the carrier is a solid. Preferably, the mixture is made into a unit dose suppository. Suitable carriers include cocoa butter and other materials commonly used in the art. Suppositories can be prepared by first forming a composition containing a softened or melted carrier, followed by cooling and shaping in a mold.

[0043] In addition to the above-mentioned carrier components, the above-mentioned drugs may include (if applicable) one or more additional carrier components, such as diluents, buffers, flavoring agents, adhesives, surfactants, thickeners, lubricants, preservatives (including antioxidants), etc. In addition, other excipients, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc., colorants and flavoring agents, etc., may be added. The preparation is made isotonic with the blood of the intended recipient. The components containing the Chinese medicine composition of the present invention can also be prepared in the form of powder or concentrate.

[0044] The medicament may also include other active ingredients.

[0045] The other active ingredients are selected from at least one of the following ingredients.

[0046] Phenytoin Sodium Valproate (VPA) Phenobarbital Lamotrigine Carbamazepine Topiramate Oxcarbazepine Levetiracetam Gabapentin Clonazepam Lacosamide Rufinamide

[0047] In a second aspect, the present invention provides use of a D-serine inhibitor in the preparation of a drug for treating a nervous system disease.

[0048] Preferably, the neurological disease has the same definition as above.

[0049] Preferably, the drug has the same definition as above.

[0050] Preferably, the D-serine inhibitor is a compound of formula I or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof, and the compound of formula I has the same definition as above.

[0051] In a third aspect, the present invention provides use of an L-serine inhibitor in the preparation of a drug for treating a nervous system disease.

[0052] Preferably, the L-serine inhibitor can inhibit the production and release of L-serine in astrocytes.

[0053] Preferably, the neurological disease has the same definition as above.

[0054] Preferably, the drug has the same definition as above.

[0055] Preferably, the L-serine inhibitor is a compound of formula I or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof, and the compound of formula I has the same definition as above.

[0056] In a fourth aspect, the present invention provides a compound or its stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug, the structural formula of which is as follows:

[0057]

[0058] Wherein, R1 has the same definition as above.

[0059] Specifically: R1 is selected from H, substituted or unsubstituted C 1-8 Alkyl, C 3-8 Cycloalkyl, aryl, heterocyclic, halogen, C 1-8 Alkoxy, the substituted substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6-membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6-membered heterocyclic group containing oxygen and / or nitrogen;

[0060] Preferably, R1 is selected from substituted or unsubstituted C 1-5 Alkyl, C 3-5 Cycloalkyl, aryl, 3-6 membered heterocyclic group, halogen, C 1-5 Alkoxy, the substituted substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6-membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6-membered heterocyclic group containing oxygen and / or nitrogen;

[0061] More preferably, R1 is selected from C 1-5 Alkyl, C 3-5 Cycloalkyl, phenyl, halogen, C 1-5 Alkoxy;

[0062] More preferably, R1 is selected from C 1-3 Alkyl, phenyl, halogen, C 1-3 Alkoxy;

[0063] More preferably, R1 is selected from C 1-3 Alkyl, halogen, C1-3 Alkoxy;

[0064] More preferably, R1 is selected from halogen, C 1-3 Alkoxy;

[0065] More preferably, R1 is selected from F, Cl, Br, C 1-3 Alkoxy;

[0066] More preferably, R1 is selected from F, methoxy;

[0067] The most preferred is F, and the preferred structural formula is as follows:

[0068]

[0069] In a fifth aspect, the present invention provides a method for preparing the compound of the fourth aspect, comprising the following steps:

[0070]

[0071] Preferably, the method specifically comprises the following steps:

[0072] (1) Add compound 1, a base, a catalyst and (Boc)2O to a solvent, and react at 20-40° C. for 1-5 hours to obtain compound 2;

[0073] (2) adding NaBH4 to the solution of compound 2 at a temperature of -10-10°C and reacting for 0.2-1 hour to obtain compound 3;

[0074] (3) Compound 3 is dissolved in a solvent, cooled to -5-5°C, and a base and MsCl (methylsulfonyl chloride) are added in sequence, and the mixture is reacted at -10-10°C for 2-4 hours to obtain compound 4;

[0075] (4) Compound 4 is added with a solvent and a reducing agent at -5-5°C under an inert atmosphere, and the mixture is heated to 20-40°C for reaction for 1-3 hours to obtain compound 5;

[0076] (5) Compound 5 is mixed with potassium cyanate and a solvent, and reacted at 55-75° C. in an inert atmosphere for 2-4 hours to obtain a product.

[0077] In a sixth aspect, the present invention provides a drug comprising the compound described in the fourth aspect or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs.

[0078] Preferably, the drug can be prepared into dosage forms such as pills, capsules, granules, oral liquids, powders, tablets, lozenges, lozenges, injections, etc., and suitable drug carriers in the art can be selected for different dosage forms.

[0079] The pharmaceutical carrier used can be solid, liquid or gas. Examples of solid carriers include lactose, kaolin, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate and stearic acid. Examples of liquid carriers include syrup, peanut oil, olive oil and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0080] When preparing drugs in oral dosage forms, any convenient pharmaceutical medium can be used. For example, water, ethanol, oil, alcohol, flavoring agents, preservatives, colorants, etc. can be used to form oral liquid preparations, such as suspensions, elixirs and solutions; while carriers, such as starch, sugars, microcrystalline cellulose, diluents, granulating agents, emulsifiers, lubricants, binders, disintegrants can be used to form oral solid preparations, such as powders, capsules and tablets. Tablets and capsules are preferred oral dosage units using solid pharmaceutical carriers due to their ease of administration. Tablets can be coated using standard aqueous or non-aqueous techniques.

[0081] Tablets can be prepared by tableting or molding, and one or more auxiliary ingredients or adjuvants can be used optionally. The active ingredient can be tableted in a free-flowing form (such as powder or granules) in a suitable machine, and can be optionally mixed with a binder, lubricant, inert diluent, surfactant or dispersant to prepare a tablet. Molded tablets can be molded in a suitable machine, that is, a powdered compound mixture moistened with an inert liquid diluent. Each tablet preferably contains about 0.05 mg to about 5 g of active ingredient, and each sachet or capsule preferably contains about 0.05 mg to about 5 g of active ingredient. For example, a preparation intended for oral administration to humans may contain about 0.5 mg to about 5 g of active drug, mixed with an appropriate and convenient carrier material, which may account for about 5% to 95% of the total composition. The unit dosage form usually contains about 1 mg to about 2 g of active ingredient, usually 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg or 1000 mg.

[0082] Drugs suitable for parenteral administration may be prepared as aqueous solutions or suspensions of the active compound. Suitable surfactants may be included, such as hydroxypropylcellulose. Dispersions may also be prepared in glycerol, liquid polyethylene glycols, and oil mixtures thereof. In addition, preservatives may be added to prevent the harmful growth of microorganisms.

[0083] The medicament of the present invention can be in a form suitable for topical use, such as an aerosol, cream, ointment, lotion, powder or the like. In addition, the composition can be in a suitable form for transdermal administration devices. For example, a cream or ointment with a desired consistency is prepared by mixing a hydrophilic material and water, and about 5wt% to about 10wt% of the compound.

[0084] The medicine of the present invention can be in a form suitable for rectal administration, wherein the carrier is a solid. Preferably, the mixture is made into a unit dose suppository. Suitable carriers include cocoa butter and other materials commonly used in the art. Suppositories can be prepared by first forming a composition containing a softened or melted carrier, followed by cooling and shaping in a mold.

[0085] In addition to the above-mentioned carrier components, the above-mentioned drugs may include (if applicable) one or more additional carrier components, such as diluents, buffers, flavoring agents, adhesives, surfactants, thickeners, lubricants, preservatives (including antioxidants), etc. In addition, other excipients, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc., colorants and flavoring agents, etc., may be added. The preparation is made isotonic with the blood of the intended recipient. The components containing the Chinese medicine composition of the present invention can also be prepared in the form of powder or concentrate.

[0086] The medicament may also include other active ingredients.

[0087] The other active ingredients are selected from at least one of the following ingredients.

[0088] Phenytoin Sodium Valproate (VPA) Phenobarbital Lamotrigine Carbamazepine Topiramate Oxcarbazepine Levetiracetam Gabapentin Clonazepam Lacosamide Rufinamide

[0089] In a seventh aspect, the present invention provides a method for treating a nervous system disease, comprising administering a specific dose of a PSPH inhibitor, a D-serine inhibitor or an L-serine inhibitor.

[0090] Preferably, the administration route medicine can be suitable for administration by any suitable route, for example, by oral (including oral or sublingual), rectal, nasal, topical (including oral, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) route administration. Such compositions can be prepared by any method known in the pharmaceutical field, for example, by mixing the active ingredient with a carrier under sterile conditions.

[0091] Further preferably, the medicament suitable for oral administration may be provided in the form of capsules or tablets; as a powder or granules; as a solution, syrup or suspension (in an aqueous or non-aqueous liquid); or as an emulsion.

[0092] Tablets or hard gelatin capsules may include lactose, corn starch or its derivatives, stearic acid or its salts.

[0093] Soft gelatin capsules may include vegetable oils, waxes, fats, semisolid or liquid polyols, and the like.

[0094] Solutions and syrups may include water, polyols and sugars. For the preparation of suspensions, oils (eg vegetable oils) may be used to provide oil-in-water or water-in-oil suspensions.

[0095] Further preferably, the drug suitable for transdermal administration can be provided as a discrete patch, the patch is used to maintain close contact with the epidermis of the recipient for a prolonged period of time. For example, the active ingredient can be delivered from the patch by iontophoresis.

[0096] Further preferably, the medicament suitable for topical administration may be provided in the form of an ointment, cream, suspension, lotion, powder, solution, paste, gel, spray, aerosol or oil.

[0097] For infections of the eye or other external tissues, such as the mouth and skin, topical ointments or creams are preferably used. When formulated as an ointment, the active ingredient may be used with a paraffinic or water-miscible ointment base. Alternatively, the active ingredient may be formulated as a cream with an oil-in-water base or a water-in-oil base.

[0098] Suitable medicaments for topical administration to the eye include eye drops. Here, the active ingredient may be dissolved or suspended in a suitable carrier, for example in an aqueous solvent.

[0099] Medicaments suitable for topical administration in the mouth include lozenges, pastilles and mouthwashes.

[0100] Further preferably, the medicament suitable for rectal administration may be provided as a suppository or an enema.

[0101] Further preferably, the medicament using a solid carrier suitable for nasal administration comprises a coarse powder (e.g. having a particle size in the range of 20-500 microns). This can be administered in the form of snuff, i.e., by rapid inhalation through the nose from a powder container close to the nose.

[0102] Compositions for nasal administration using liquid carriers include nasal sprays or nose drops. These may include aqueous or oily solutions of the active ingredient.

[0103] Medicaments suitable for administration by inhalation include fine particle dusts or mists which may be generated by various types of devices, such as pressurized aerosols, nebulizers or sprays. Such devices may be constructed to provide a predetermined dose of the active ingredient.

[0104] Further preferably, the medicament suitable for vaginal administration may be provided as a vaginal suppository, tampon, cream, gel, paste, foam or spray formulation.

[0105] Further preferably, the medicine suitable for parenteral administration includes aqueous and non-aqueous sterile injectable solutions or suspensions. These compositions may include antioxidants, buffers, antibacterial agents and solutes to make the compositions substantially isotonic with the blood of the intended recipient. Other components that may be present in these compositions include, for example, water, alcohol, polyols, glycerol and vegetable oils. Compositions suitable for parenteral administration may be present in unit doses or multidose containers, such as sealed ampoules and vials, and may be stored immediately before use in freeze-dried (lyophilized) conditions, which only require the addition of sterile liquid carriers, such as sterile water for injection. Instant injection solutions and suspensions can be prepared from sterile powders, granules and tablets.

[0106] Preferably. The dosage is easily determined by routine experimentation and controlled by a physician or clinician. The guiding principle for determining an appropriate dosage is to deliver an amount of material that is appropriately effective but non-toxic or of acceptable toxicity. For NB-DNJ or similar compounds, the daily dosage for an adult can be expected to be in the range of 1 mg-2 g of active agent, and may be in the range of 100-800 mg or 300-600 mg. The dosage may be administered in a single dose in a day, or in two, three or more doses in a day.

[0107] Preferably, the PSPH inhibitor, D-serine inhibitor or L-serine inhibitor is administered in combination with other active ingredients.

[0108] The other active ingredients are selected from at least one of the following ingredients.

[0109] Phenytoin Sodium Valproate (VPA) Phenobarbital Lamotrigine Carbamazepine Topiramate Oxcarbazepine Levetiracetam Gabapentin Clonazepam Lacosamide Rufinamide

[0110] Preferably, the PSPH inhibitor, D-serine inhibitor or L-serine inhibitor has the same definition as above.

[0111] Explanation and explanation of terms:

[0112] In the present invention, the term "prevention" refers to a method of allowing a subject to avoid or circumvent a disease or condition or to postpone the recurrence or appearance of one or more symptoms of the condition. The term "improvement" refers to a method of alleviating symptoms that have already occurred in a subject or slowing down the trend of symptom development. The term "treatment" refers to obtaining the desired pharmacological and / or physiological effect, which can be preventive in terms of completely or partially preventing a disease or its symptoms, and / or can be therapeutic in terms of partially or completely curing a disease and / or the adverse effects caused by the disease. The term "treatment" as used herein includes any treatment of a disease in mammals, especially humans, and includes (a) preventing the occurrence of the disease in a subject who is susceptible to a disease but has not yet been diagnosed with the disease; (b) inhibiting the disease, that is, preventing its development; and (c) eliminating the disease, even if the disease subsides and / or eliminates one or more disease symptoms. "Treatment" should also include the delivery of an agent to provide a pharmacological effect, even if the disease or condition does not exist.

[0113] In the present invention, the terms “comprise”, “contain” and “include” mean including but not limited to, and do not exclude other additives and components, for example.

[0114] Unless otherwise specified, the term "alkyl" as used herein includes branched and straight chain saturated aliphatic hydrocarbon groups having a specified number of carbon atoms, including all isomers. Common abbreviations for alkyl groups, such as methyl, can be represented by "Me" or CH3, ethyl, can be represented by "Et" or CH2CH3, propyl, can be represented by "Pr" or CH2CH2CH3, butyl, can be represented by "Bu" or CH2CH2CH2CH3, etc. For example, "C 1-4 "C1-C4 alkyl" refers to a straight or branched chain alkyl group having a specified number of carbon atoms, including all isomers. 1-4 Alkyl includes n-, iso-, sec- and tert-butyl, n- and iso-propyl, ethyl and methyl. 1-10 "Alkyl" and the like have similar meanings.

[0115] The term "alkoxy" refers to straight and branched chain alkyl groups having the indicated number of carbon atoms attached through an oxygen bridge.

[0116] The term "halogen" (or halo) refers to fluorine, chlorine, bromine and iodine (alternatively referred to as fluoro (F), chloro (Cl), bromo (Br) and iodo (I)).

[0117] The term "aryl" refers to aromatic mono- and polycyclic carbon ring systems wherein the individual carbon rings in the polycyclic system are fused or linked to each other by single bonds. Typical aryl groups include phenyl, naphthyl and biphenylene.

[0118] The term "heterocycle" refers to a ring structure composed of carbon atoms and non-carbon atoms, such as nitrogen, oxygen and sulfur. Typical heterocyclic groups include pyridine, quinoline, tropane, phenothiazine, benzodiazepine, furan, pyrazolone and pyrimidine.

[0119] The term "aromatic heterocycle" refers to a 5- or 6-membered monocyclic aromatic ring or a 7- to 12-membered bicyclic ring consisting of carbon atoms and one or more heteroatoms selected from N, O and S. Examples of aromatic heterocycles include pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl (or thienyl), thiazolyl, furanyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl and thiadiazolyl, benzotriazolyl, indolyl, isoindolyl, indazolyl, indolinyl, isoindolinyl, quinoxalinyl, quinazolinyl, cinnolinyl, chromanyl, isochromanyl, tetrahydroquinolyl, quinolyl, tetrahydroisoquinolyl, isoquinolyl, 2,3-dihydrobenzofuranyl, 2,3-dihydrobenzo-1,4-dienyl, imidazo(2,1-b)(1,3)thiazole and benzo-1,3-dioxolyl.

[0120] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid. When the compound of the present invention is acidic, its corresponding salt can be easily prepared from an inorganic base or an organic base. Salts derived from such inorganic bases include salts of aluminum, ammonium, calcium, copper (copper and cuprous), iron, ferrous, lithium, magnesium, manganese (manganese and manganous), potassium, sodium, zinc, etc. Preferred are salts of ammonium, calcium, magnesium, potassium and sodium. Salts prepared from organic bases include primary, secondary and tertiary amines derived from natural and synthetic sources. Pharmaceutically acceptable organic non-toxic bases that can form salts include arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydramine, isopropylamine, dicyclohexylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When the compound of the present invention is basic, its corresponding salt can be easily prepared from inorganic or organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, and the like.

[0121] The term "solvate" refers to a complex of variable stoichiometry formed by a solute (i.e., a compound of Formula I) or a pharmaceutically acceptable salt thereof and a solvent that does not interfere with the biological activity of the solute. Examples of solvents include, but are not limited to, water, ethanol, and acetic acid. When the solvent is water, the solvate is referred to as a hydrate. Hydrates include, but are not limited to, hemihydrates, monohydrates, one and a half hydrates, dihydrates, and trihydrates.

[0122] The term "prodrug" is a functional derivative of the compounds of the invention which is readily convertible in vivo into the desired compound.

[0123] The beneficial effects of the present invention are:

[0124] The present invention finds that PSPH (phosphoserine phosphatase) can be used as a new therapeutic target for nervous system diseases, especially epilepsy, thereby providing a new direction for the treatment of nervous system diseases. At the same time, based on the target of PSPH, the present invention provides a small molecule compound that can be used as a PSPH inhibitor, and uses a small molecule chemical inhibitor to inhibit the activity of PSPH to achieve the treatment of nervous system diseases, especially to exert an excellent anti-epileptic effect. The newly discovered PSPH inhibitor compound shows good pharmacological properties, exhibits good anti-epileptic effects in the mouse TLE model, and has less toxic side effects than other channel blockers targeting NMDARs. BRIEF DESCRIPTION OF THE DRAWINGS

[0125] Figure 1 Figure 1 shows that D-serine promotes KA-induced acute epileptic seizures. (ac) Metabolomics analysis of glutamate, serine, and glycine levels in hippocampal samples from patients with hippocampal sclerosis (HS, n = 6), non-hippocampal sclerosis (nonHS, n = 3), and autopsy controls without neurological diseases (n = 7). (d, e) Microdialysis and zero-flow regression were used to measure hippocampal L / D-serine levels in control mice and chronic TLE mice, and statistical analysis was performed (n = 6). (f) Representative 2-minute EEG schematic. The kainate-induced acute epilepsy model was established, and low-dose D-serine (10 μM), high-dose D-serine (100 μM), D-amino acid oxidase (1 U / mL), heat-inactivated amino acid oxidase (1 U / mL), 7-CKA (100 μM), or glycine (200 μM) were injected into the epileptic focus in advance. (gk) Differences in the onset time of epileptic seizures (g), total number of epileptic seizures (h), time of each epileptic seizure (i), total number of electroclinical epileptic seizures (j) and epileptic inducibility rate (k) among the control group, low-dose D-serine group, high-dose D-serine group, D-amino acid oxidase group, hiDAAO group, 7-CKA group and glycine group were analyzed (n=9).

[0126] Figure 2Figure 1 shows the regulation of spontaneous epileptic seizures by D-serine. (a) Schematic diagram of the experimental scheme (glycine group n = 4, other groups n = 8). (bp) Representative 1-minute EEG of each group (left), schematic diagram of the distribution of epileptic seizures during the experimental process (middle), and statistical analysis of hippocampal paroxysmal discharges at baseline, 0-12 hours after injection, and 12-28 hours after injection after unilateral hippocampal injection of solvent (d), D-amino acid oxidase (g), 7-CKA (j), D-serine (m), or glycine (p) (right). After 20 hours of EEG baseline detection, 0.5 μL of solvent, D-amino acid oxidase (1 U / ml), 7-CKA (100 μM), D-serine (10 μM), or glycine (200 μM) was injected into the ipsilateral hippocampus of kainate injection at 8:00 on the second day through the pre-implanted catheter to observe the changes in hippocampal paroxysmal discharges.

[0127] Figure 3 Figure 1. L-serine produced by astrocytes regulates D-serine levels and epileptic seizures. (a) Schematic diagram of the serine shuttling pathway between astrocytes and neurons. (b) Schematic diagram of the reverse microdialysis (1 μL / min) and simultaneous EEG recording protocol. (c) Schematic diagram of the experimental process and group information. (d) Schematic diagram of hippocampal paroxysmal discharges induced by reverse microdialysis injection of KA into the hippocampus. (ej) Raster plots showing the effect of specific treatments on hippocampal paroxysmal discharges during epileptic seizures, with each occurrence of hippocampal paroxysmal discharges indicated by a red line. (km) Statistical analysis of the number of epileptic seizures (k) and the levels of L-serine (l) and D-serine (m) in the dialysate (n = 6).

[0128] Figure 4Figure 2 shows that PSPH is upregulated in TLE and regulates interstitial L-serine and D-serine levels. (a) Schematic diagram of the three-step enzymatic reaction for de novo synthesis of L-serine by astrocytes. (b, c) PSPH, PSAT1, or PHGDH in astrocytes were knocked down using a set of siRNAs, and L-serine levels in cell lysates (b) and culture supernatants (c) were statistically analyzed. (d) PSPH, PSAT1, and PHGDH levels in seven different brain regions of mice were detected (left) and statistically analyzed (right). (e) Immunoblotting (left) and statistical analysis (right) of mouse hippocampal lysates 0, 1, 2, and 4 h after intraperitoneal injection of pentylenetetrazol (PTZ) or PTZ+chloral hydrate (CH). (f, g) Immunofluorescence staining of GFAP and PSPH in the hippocampus of surgically resected temporal lobes of patients with hippocampal sclerosis (TLE-HS) and autopsied controls without neurological disease (control) (f) or in the hippocampus of normal mouse controls and mice with temporal lobe epilepsy (4 weeks after kainate-induced status epilepticus) (g); scale bar = 50 μm. (h) Immunofluorescence staining of PSPH, GFP, and GFAP in the hippocampus of mice one month after bilateral hippocampal injection of AAV5-gfa104-eGFP (control) or AAV5-gfa104-PSPH-IRES-eGFP (PSPH overexpression); scale bar = 50 μm. (i, j) Levels of L-serine (i) and D-serine (j) in the extracellular fluid of the hippocampus of control and PSPH overexpressing mice were measured using microdialysis (n = 6). (kn) Acute epileptic seizures were induced by injecting kainate (7 ng) into the hippocampus of control and PSPH-overexpressing mice through a pre-implanted catheter. The differences in the responses of control and PSPH-overexpressing mice to kainate, including the onset time of epileptic seizures (k), the number of epileptic seizures (l), the duration of each epileptic seizure (m), and the number of electroclinical seizures (n) were statistically analyzed (n=8).

[0129] Figure 5 This is the mass spectrum analysis diagram of compound 1.

[0130] Figure 6 This is the mass spectrum analysis diagram of compound 2.

[0131] Figure 7 This is the mass spectrum analysis diagram of compound 3.

[0132] Figure 8The results of the PSPH inhibitory activity test of compound 1. (a, b) The content of L-serine in the supernatant and cell lysate. Primary astrocytes treated with compound 1 were treated with compound 1 for 24 hours in serine-free medium, and then the medium was replaced with fresh serine-free medium. The cells were cultured for another 24 hours, and the levels of L-serine in the supernatant and cell lysate were detected (n = 4). (c, d) After chronic epileptic mice were intraperitoneally injected with solvent or 4 mg / kg Z218484536 five times, the levels of L-serine (t) and D-serine (u) in the hippocampus were measured by microdialysis (n = 6).

[0133] Fig. 9 The results of the PSPH inhibitory activity test of compound 2. (a) In an in vitro test tube, different concentrations of compound 2 were added to observe the inhibitory ability of recombinant PSPH to degrade the substrate O-phospho-L-serine to produce L-serine. (b, c) The content of L-serine in the supernatant and cell lysate. Primary astrocytes treated with compound 2 were treated with 40-2000μM compound 2 in serine-free medium for 24 hours, and then fresh serine-free medium was replaced. The cells were cultured for another 24 hours, and the levels of L-serine in the supernatant and cell lysate were detected (n=4).

[0134] Fig.10 The results of the PSPH inhibitory activity test of compound 3. (a) In an in vitro test tube, different concentrations of derivative-2 were added to observe the inhibitory ability of recombinant PSPH to degrade the substrate O-phospho-L-serine to produce L-serine. (b, c) The content of L-serine in the supernatant and cell lysate. Primary astrocytes treated with compound 3 were treated with compound 3 in a serine-free medium for 24 hours, and then the medium was replaced with fresh serine-free medium, and the cells were cultured for another 24 hours, and the level of L-serine in the supernatant and cell lysate was detected (n=4).

[0135] Fig.11 The following are the animal test results of four compounds.

[0136] Fig.12 Schematic diagram of the molecular docking experiment and pharmacokinetics of compound 1.

[0137] Fig.13Schematic diagram of the target action of compound 1. (a) PSPH protein sequence alignment among 11 species revealed that the four binding sites of PSPH are highly evolutionarily conserved. (b) Enzyme activity assay of wild type (WT) and PSPH mutants with mutations at Asp22Ala, Ala51Val, Ala51Gly, Gly110Val, Gly110Ala, Lys158Ala or Ala51Val / Gly110Ala sites. (c) MST analysis of compound 1 binding to PSPH Ala51Val / Gly110Ala mutant. (d, e) L-serine content in cell lysate (d) and culture supernatant (e). PSPH knockdown and PSPH-MT overexpression (Ala51Val / Gly110Ala) were achieved by lentivirus-mediated gene transfer.

[0138] Figure 14-15 Activity test of compound 1 on a panel of 25 protein phosphatases. Statistical analysis of the inhibitory effects of (az)Z218484536 and positive controls on PSPH, 20 protein tyrosine phosphatase superfamily enzymes (including SHP1, SHP2, PTPN2, PTBN4, PTPN7, PTPN9, PTPN12, PTPN13, PTPN22, PTPRB, PTPRC, PTPRE, PTPRM, DUSP3, DUSP10, DUSP13, DUSP22, YopH, LMPTP-a, LMPTP-B) and five protein serine / threonine phosphatase superfamily enzymes (including PP1A, PP1B, PP2A, λPP and PP5). The left side of each panel is the known positive inhibitor of the enzyme, and the right side is the test result of compound 1.

[0139] Fig.16 Compound 1 exhibits antiepileptic effect in the kainic acid chronic temporal lobe epilepsy mouse model. (a) Experimental design scheme. (be) The number of spontaneous convulsive seizures in the solvent, lamotrigine and Z218484536 groups was recorded daily. Lamotrigine was administered intraperitoneally at a dose of 20 mg / kg per day (n=8). Z218484536 was administered intraperitoneally at a dose of 2 or 4 mg / kg per day (n=9). DMSO was used as a solvent control (n=8). (f) The line graph shows the daily seizure frequency of mice during the 3-week baseline recording and the subsequent 7-week drug treatment period. (gj) Statistical analysis of the seizure frequency of the solvent group, lamotrigine group, compound 1 dose 2 mg / kg group and 4 mg / kg group before and after treatment. DETAILED DESCRIPTION

[0140] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the invention of the present invention based on the disclosed content, and they should also fall within the scope of the present application.

[0141] The present invention is further described below by way of specific examples. The various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the contents described below are all weight contents. Unless otherwise specified, it is understood that the process is carried out at room temperature.

[0142] In the following examples, the meanings of the abbreviations are as follows: PSPH: Phosphoserine Phosphatase; PSAT1: Phosphoserine Aminotransferase 1; PHGDH: phosphoglycerate dehydrogenase; ASM: Anti-seizure Medication; NMDAR: N-methyl-D-aspartate-receptor; SR: Serine Racemase; TLE: temporal lobe epilepsy; HS: hippocampal sclerosis; NonHS: non hippocampal sclerosis; KA: Kainic acid; DAAO: D-α-amino acid oxidase; hiDAAO: Heatinactivated D-α-amino acid oxidase, heat-inactivated DAAO; 7-CKA: 7-Chlorokynurenic acid, 7-chlorokynurenic acid; Gly: Glycine, glycine; HPDs: hippocampal paroxysmal discharges, hippocampal paroxysmal discharges; CH: chloral hydrate, chloral hydrate; SE: status epilepticus, status epilepticus; GFP: Green fluorescent protein, green fluorescent protein; GFAP: glial fibrillary acidic protein, glial fibrillary acidic protein; HTVS: High-throughput virtual screening, high-throughput virtual screening; PTZ: pentylenetetrazol, pentylenetetrazol; PK: Pharmacokinetics, pharmacokinetics; CCK8: Cell CountingKit-8, cell viability detection; LD50: Lethal Dose, 50%, half lethal dose; HepG2: Human hepatocellular carcinomas, liver cancer cells; LTG: lamotrigine, lamotrigine; AST: Aspartateaminotransferase, aspartate aminotransferase; ALT: Alanine aminotransferase, alanine aminotransferase;BUN: blood urea nitrogen; Cr: creatinine; ASCT1 / 2: Alanine / Serine / Cysteine / Threonine Transporter1 / 2; ASC1: Alanine / Serine / Cysteine ​​Transporter1; L-4FPG: L-4-fluorophenylglycine; ATA1 / 2: amino acid transporter1 / 2; Gln: Glutamine; KO: Knockout; OE: Overexpression; SESs: Spontaneous electroclinical seizures; IC50: half maximal inhibitory concentration; Kd: The equilibrium dissociation constant; IP: Intraperitoneal Injections, intraperitoneal injection; LC-MS: Liquid Chromatograph Mass Spectrometer; EEG: Electroencephalogram; NAD: Nicotinamide Adenine Dinucleotide; KEGG: Kyoto Encyclopedia of Genes and Genomes; RT (rt): room temperature, usually 25°C.

[0143] Example 1: Verification experiment of PSPH as a therapeutic target

[0144] (1) Effects of D-serine on acute epileptic seizures induced by kainate (KA)

[0145] The levels of neurotransmitters were analyzed in hippocampal samples from patients with hippocampal sclerosis, patients without hippocampal sclerosis, and autopsy controls without neurological disease. Figure 1 As shown in a and b, the concentrations of glutamate and serine in hippocampal sclerosis samples were significantly increased compared with normal samples and non-hippocampal sclerosis samples, while there was no significant difference in the level of glycine among the three groups ( Figure 1c). To further detect the levels of L-serine and D-serine in the extracellular fluid of the hippocampus, a kainic acid-induced chronic epilepsy mouse model was established. Four weeks after unilateral hippocampal injection of kainic acid, the mice developed spontaneous recurrent epileptic seizures. Through microdialysis and zero-flow back-calculation, we calculated that the levels of L-serine and D-serine in the control group of mice injected with normal saline were 7.1μM and 1.5μM, respectively ( Figure 1 d, e in the figure). After the injection of kainic acid, the levels of L-serine and D-serine in the extracellular fluid of the mouse hippocampus increased by 2.7 times (19.0 μM) and 3.7 times (5.5 μM), respectively, compared with the control group. Excessive D-serine is released into the postsynaptic membrane and acts on the NMDAR receptor to further regulate synaptic activity, thereby affecting epileptic seizures, indicating that changes in D-serine levels may affect kainic acid-induced acute epileptic seizures. Unilateral injection of 7 ng of kainic acid into the hippocampus of awake mice through a pre-implanted catheter can induce several hippocampal paroxysmal discharges (HPDs) within about an hour, and a few mice will show convulsion symptoms, which appear as electroclinical seizures in EEG recordings ( Figure 1 f), it was found that pre-injection of low-dose D-serine (10 μM) could significantly shorten the onset time of KA-induced epileptic seizures ( Figure 1 g in ), and lead to a more severe epileptic phenotype, including an increase in the number of seizures ( Figure 1 h in), the increase in the duration of each epileptic seizure ( Figure 1 i) and an increase in the number of electroclinical seizures ( Figure 1 In contrast, pre-injection of D-amino acid oxidase (DAAO) to degrade extracellular D-serine almost completely suppressed kainate-induced acute epileptic seizures ( Figure 1 h, j, k in the figure). We heated D-amino acid oxidase at 95℃ for 3 minutes to inactivate it, and then injected the heat-inactivated D-amino acid oxidase into the mouse brain, and found that it did not inhibit the epileptic seizures of the mice ( Figure 1 In addition, 7-CKA (7-chlorokynurenic acid), a selective antagonist of the glycine site of NMDARs, can also significantly inhibit KA-induced acute epileptic seizures, but we speculate that this effect is not caused by glycine because pre-injection of glycine has no proconvulsant effect ( Figure 1 gk in the hippocampus). It was also found that the effect of high-dose D-serine injected into the hippocampus was opposite to that of low-dose D-serine, and epileptic seizures were suppressed under the action of high-dose serine ( Figure 1This inhibitory effect may be due to the nonspecific competitive binding of high-dose D-serine to the glutamate binding site of NMDARs. In summary, D-serine promotes KA-induced acute epileptic seizures.

[0146] (2) D-serine regulates spontaneous recurrent epileptic seizures

[0147] To further demonstrate the effect of D-serine on spontaneous chronic epileptic seizures, we established a mouse model of chronic temporal lobe epilepsy. Four weeks after unilateral injection of kainic acid into the hippocampus of mice, electrodes were implanted under the skull for EEG monitoring and catheters were implanted for hippocampal drug injection ( Figure 2 We first recorded the EEG of mice with temporal lobe epilepsy for 20 hours to obtain the baseline of hippocampal paroxysmal discharges. Compared with the baseline ( Figure 2 bd in the hippocampus), D-amino acid oxidase can significantly inhibit hippocampal paroxysmal discharges within 12 hours after hippocampal injection ( Figure 2 The same results were observed when mice were injected with 7-CKA ( Figure 2 However, injection of additional D-serine did not increase the number of hippocampal paroxysmal discharges ( Figure 2 km), we speculate that in mice with temporal lobe epilepsy, D-serine on NMDARs may have been saturated, and injection of glycine has no effect on hippocampal paroxysmal discharges ( Figure 2 Together, these data suggest that upregulation of D-serine is a prerequisite for the development of epileptic seizures in mice with chronic temporal lobe epilepsy, and that eliminating D-serine or blocking its effects on NMDARs can exert anti-epileptic effects.

[0148] (3) L-serine produced by astrocytes can regulate D-serine levels and epileptic seizures

[0149] Astrocytes are the main cell type that synthesizes L-serine in the brain. They synthesize L-serine from glucose through a three-step enzymatic reaction, which is then released and taken up by surrounding neurons, where it is converted into D-serine by the action of serine racemase. This process is called "serine shuttle." Therefore, we tried to prove whether L-serine produced by astrocytes would affect D-serine levels and epileptic seizures. Figure 3As shown in a, L-serine is first released by astrocytes through alanine / serine / cysteine / threonine transporters 1 and 2 (ASCT1 / 2), then taken up by amino acid transporters 1 and 2 (ATA1 / 2) into neurons and converted into D-serine under the action of serine racemase, and finally released through neuronal alanine serine cysteine ​​transporter-1 (Asc-1) to promote the activation of synaptic NMDARs. To verify the role of the above pathway in epilepsy, we injected kainate and inhibitors of the above transporters into the hippocampus of awake mice by reverse microdialysis, synchronously collected the dialysate to analyze its serine level, and detected epileptic seizures in real time by electroencephalography (EEG). Figure 3 b) The experiment set up 6 groups in total, and the group settings were as follows Figure 3 As shown in c. We first perfused 7ng / μL kainate at a rate of 1μL / min and found that epilepsy could be successfully induced, as shown in EEG as hippocampal paroxysmal discharges ( Figure 3 d). Compared with the solvent control group, 1-hour perfusion of kainate induced 4 to 5 hippocampal paroxysmal discharges ( Figure 3 When L-4FPG was infused to inhibit ASCT1 and ASCT2, KA-induced epileptic seizures were almost completely blocked ( Figure 3 g in the figure). When exogenous L-serine is supplemented again, it can offset the inhibitory effect of L-4FPG on hippocampal paroxysmal discharges ( Figure 3 When we further inhibited neuronal L-serine uptake with glutamine (inhibiting ATA1 and ATA2) or blocked neuronal D-serine release with BMS-466442 (inhibiting Asc-1), hippocampal paroxysmal firing was almost abolished ( Figure 3 The statistical results of epileptic seizures in the above groups are detailed in Figure 3 We also tested the levels of L-serine and D-serine in the dialysate of each group and found that L-4FPG effectively prevented the increase of L-serine and D-serine levels induced by kainate ( Figure 3 l, m in the figure). In the presence of L-4FPG, exogenous L-serine will again cause an increase in D-serine; glutamine or BMS-466442 will further block the increase in D-serine ( Figure 3 In conclusion, L-serine produced by astrocytes plays an important role in regulating interstitial D-serine levels and epileptic seizures.

[0150] (4) PSPH is a key target for regulating L-serine production and release in astrocytes

[0151] Given the potential role of L-serine in modulating epileptic seizures, we examined whether modulation of L-serine synthesis could be an effective anti-epileptic approach. Figure 4 As shown in a, astrocytes synthesize L-serine from the glycolytic intermediate 3-phosphoglycerate through a three-step enzymatic reaction, which is sequentially catalyzed by phosphoglycerate dehydrogenase (PHGDH), phosphoserine aminotransferase 1 (PSAT1), and phosphoserine phosphatase (PSPH). PSPH is a therapeutic target based on the following evidence. First, compared with PSAT1 and PHGDH, the knockdown of PSPH resulted in the most significant decrease in intracellular and extracellular L-serine levels ( Figure 4 Second, we measured the expression of these three enzymes in seven different brain regions of mice and found that the PSPH content was highest in the hippocampus ( Figure 4 d in the figure), and the hippocampus is the epileptogenic brain region of temporal lobe epilepsy. In addition, we also found that the expression of PSPH protein in the hippocampus is related to neuronal activity ( Figure 4 Finally, the level of PSPH was abnormally increased in the hippocampus of epileptogenic lesions surgically removed from patients with temporal lobe epilepsy and in the hippocampal astrocytes of mice with chronic temporal lobe epilepsy ( Figure 4 Through the above experiments, we speculated that regulating PSPH levels would affect the level of serine and epileptic seizure activity in vivo. To confirm this hypothesis, we first overexpressed PSPH in hippocampal astrocytes of normal mice using AAV5-gfa104-PSPH-IRES-eGFP ( Figure 4 h), and used micropermeabilization to detect the levels of L-serine and D-serine in the hippocampus, and found that both were significantly increased ( Figure 4 i, j in the figure). Compared with the control mice, PSPH-overexpressing mice were more susceptible to acute epileptic seizures induced by kainate, as shown by a shorter onset time, more frequent seizures, longer duration of each seizure, and more electroclinical seizures after kainate injection. Figure 4 kn in ).

[0152] Example 2 Compound Synthesis Example

[0153] (1) Synthesis of Compound 1

[0154]

[0155] In a solution of 20 mL of 50% alcohol, hydrazinecarboxamide hydrochloride (0.59 g, 5.26 mmol) was added to 3-(4-fluorophenyl)-1H-pyrazole-4-carbaldehyde (1 g, 5.26 mmol), and then refluxed for 1 hour. After cooling, the precipitate was filtered and recrystallized in ethanol to obtain a white solid product (600 mg, 46.15%).

[0156] LCMS(5_95_3min,Rt=1.540min),MS(ESI):m / z=248.0[M+H]+1H NMR (400MHz, DMSO-d6): δ13.26,13.19(s,1H),9.91(s,1H),8.27,8.02(s,1H) ,7.91,7.87(s,1H),7.60–7.58(m,2H),7.40–7.26(m,2H),6.30-6.26(m,2H).

[0157] (2) Synthesis of Compound 2

[0158]

[0159] To a solution of 3-(4-methoxyphenyl)-1H-pyrazole-4-carbaldehyde (200 mg, 0.99 mmol) in 4.0 mL of 50% ethanol, hydrazinecarboxamide hydrochloride (111 mg, 0.99 mmol) was added, and the reaction mixture was refluxed for 1 hour. After cooling, the precipitate was filtered and recrystallized from 50% ethanol to give the product (150 mg, 58.43% yield) as a light yellow solid.

[0160] LCMS:IDSUF06-DE-2-4R-GU Method:5_95_3min,t=1.509min,MS(ESI):m / z=260.0[M+H]

[0161] 1H NMR (400.30MHz, DMSO) δ13.13 (s, 1H), 9.88 (s, 1H), 8.11 (d, J = 92.9Hz, 1H), 7.89(s,1H),7.47(d,J=7.8Hz,2H),7.06(s,2H),6.29(s,2H),3.81(s,3H).

[0162] (3) Synthesis of Compound 3

[0163]

[0164] Step 1. Add 3-(4-fluorophenyl)-1h-pyrazole-4-carboxylic acid (5.0 g, 26 mmol) solution to THF (50 mL), add TEA (7.87 g, 78 mmol), DMAP (0.3 g, 2.6 mmol) and (Boc)2O (17.0 g, 78 mmol). The mixture was stirred at room temperature for 3 hours. The resulting mixture was diluted with water (300 mL) and extracted with ethyl acetate (3×200 mL). The combined organic layers were washed with brine (200 mL×3) and dried over anhydrous Na2SO4. The filtered filtrate was concentrated under vacuum. The crude product was purified by silica gel column eluted with EA / PE (0-10%) to obtain tert-butyl 3-(4-fluorophenyl)-4-formyl-1h-pyrazole-1-carboxylate (7.0 g, 24.11 mmol, 92%) as a white solid.

[0165] LCMS: m / z calcd. ([M+H-56]+):235.0

[0166] Step 2. Add NaBH4 (1.9 g, 52 mmol) to a solution of tert-butyl 3-(4-fluorophenyl)-4-formyl-1h-pyrazole-1-carboxylate (5.0 g, 26 mmol) in MeOH (50 mL) at 0°C. Stir the mixture at 0°C for 0.5 hours. Dilute the resulting mixture with water (300 mL). Extract the resulting mixture with ethyl acetate (3×300 mL). The combined organic layers were washed with brine (300 mL×3) and dried over anhydrous Na2SO4. The filtered filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography and eluted with [PE / EtOAc] (10:1) to give the desired product (3.5 g, 11.97 mmol, 70%) as a white solid.

[0167] LCMS: m / z calcd.([M+H]+):237.0

[0168] Step 3: Dissolve tert-butyl 3-(4-fluorophenyl)-4-(hydroxymethyl)-1h-pyrazole-1-carboxylate (0.50 g, 1.71 mmol) in DCM (5 mL) and cool to 0°C. Add TEA (865.46 mg, 8.55 mmol) and MsCl (293.89 mg, 2.57 mmol) in turn and stir at 0°C for 3 hours. Concentrate the mixture by evaporator, add water, extract with chloroform, dry and concentrate over magnesium sulfate to obtain a light orange oily compound.

[0169] To the compound solution obtained by stirring in DMF (10 mL), NaCN (125.74 mg, 2.59 mmol) was added at RT, and the reaction mixture was stirred at RT for 3 h. The reaction process was monitored by TLC. The reaction mixture was poured into ice water and extracted with ethyl acetate (2×200 mL). The combined organic layer was washed with brine (200 mL), dried over anhydrous Na2S04, filtered and concentrated under reduced pressure to obtain the title compound (250 mg, 1.24 mmol, 72%) as a light yellow oil.

[0170] LCMS:m / z calcd.([M+H+ACN]+):243.0

[0171] Step 4, 2-(3-(4-fluorophenyl)-1h-pyrazol-4-yl)acetonitrile (250mg, 1.24mmol) was added to THF (5mL) solution under nitrogen at 0°C, and then LiAlH4 (2mL, 2mmol, 1M / THF) was added, and the mixture was heated to room temperature and stirred for 2 hours. After quenching with 5mL of water, the residue was extracted with ethyl acetate (20mL). The organic layer was washed with saturated saline solution (5mL*2). The organic layer was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated in vacuo and purified by tlc (5% methanol / dichloromethane) to obtain 2-(3-(4-fluorophenyl)-1h-pyrazol-4-yl)ethylamine (120mg, 584.69umol, 47%) as a yellow solid.

[0172] LCMS: m / z calcd.([M+H]+):206.2

[0173] Step 5. Mix 2-(3-(4-fluorophenyl)-1h-pyrazol-4-yl)ethylenediamine (110 mg, 0.54 mmol) and potassium cyanate (48.19 mg, 0.59 mmol) with methanol (5 mL) and AcOH (0.2 mL), and stir under nitrogen at 65°C for 3 hours. After filtering the mixture, the filtrate was concentrated in vacuo and purified by pre-HPLC (10 mmol / L NH4HCO3 in H2O, B:ACN) to obtain 1-(2-(3-(4-fluorophenyl)-1h-pyrazol-4-yl)ethyl)urea (21 mg, 84.59 umol, 15.78%) as a white solid.

[0174] LCMS: m / z calcd.([M+H]+):249.2

[0175]

[0176] Example 4 PSPH inhibitory activity test

[0177] Test method: PSPH-FLAG recombinant protein (recombinant PSPH) was expressed and purified in HEK293 cells, the substrate O-phospho-L-serine was hydrolyzed to L-serine in an in vitro reaction system, and the L-serine level in the test tube was detected by LC-MS. Each inhibitor was pre-mixed with PSPH before the reaction to test the inhibitor effect.

[0178] The test results are as follows Figure 8-Figure 10 shown.

[0179] Figure 8 The results showed that compound 1 inhibited the release of L-serine from astrocytes into the culture medium with a half-maximal inhibitory concentration (IC50) of 0.4 μM, and inhibited intracellular L-serine levels with an IC50 of 0.38 μM. The serine level in the extracellular fluid of the hippocampus was analyzed using a kainate mouse chronic temporal lobe epilepsy model combined with microdialysis technology. The results showed that after three consecutive intraperitoneal injections of compound 1, the levels of L-serine and D-serine in the hippocampus were significantly reduced. These data indicate that compound 1, which can penetrate into the brain, has a relatively strong binding affinity with PSPH, is less toxic, and has anti-epileptic potential.

[0180] Fig. 9 The results showed that some key atoms in the structure of Z218484536 interact with PSPH. We considered whether the fluorine atom at position 16 on the benzene ring could be replaced. Therefore, we synthesized a derivative, 16-Methoxy-Z218484536, which replaced the original fluorine atom with a methoxy group. We found that this change in chemical structure did not change the ability to inhibit the activity of recombinant PSPH protease in vitro ( Fig.10 a), nor did it change the inhibitory capacity on the production and release of primary astrocytes ( Fig.10 b, c). This indicates that some substitutions of the 16-position group will not change the inhibitory ability of the compound to PSPH.

[0181] Fig.10 The results showed that the C=N bond was changed to C—C bond at position 5-6. It was found that it also has the ability to inhibit PSPH enzyme activity and inhibit the production and release of L-serine in primary astrocytes.

[0182] Example 5 Anti-epileptic effect verification

[0183] Test method: The acute mouse epilepsy model induced by pentylenetetrazol (PTZ) was used. Each compound was given to mice by intraperitoneal injection for 3 consecutive days at a dose of 0.5, 1, 2, 4, and 20 mg / kg. The starting time of PTZ-induced epilepsy and the racine seizure level of epilepsy were evaluated.

[0184] The results showed that compound 1 had significant anti-epileptic effects, including prolonging the onset time of PTZ-induced epileptic seizures and reducing the severity of epileptic seizures. Its anti-epileptic effect reached the highest level at a dose of 4 mg / kg ( Fig.11 ac in the above formula), where Z218484536 is compound 1. At the same time, we tested the effects of some close compounds, and the results are as follows Fig.11 As shown in dl, the other three compounds have no anti-epileptic effect.

[0185] Example 6 Molecular docking experiment and pharmacokinetics

[0186] The pyrazole NH group of compound 1 forms a hydrogen bond with Ala51 of PSPH, and the urea group forms four hydrogen bonds with Asp22, Gly110 and Lys158 of PSPH. In addition, the benzene ring of compound 1 forms a cation-π interaction with Lys158. The results of microcalorimetric surge experiments showed that the dissociation constant (Kd) of PSPH binding to compound 1 was about 0.27μM, indicating that the intermolecular attraction was relatively strong. The competition study of compound 1 with O-phospho-L-serine showed the competitive inhibition mode of compound 1 on PSPH.

[0187] In vivo evaluation of the pharmacokinetics (PK) of compound 1 in mice showed that after intraperitoneal injection at a dose of 4 mg / kg, the compound had good exposure (AUClast = 1088h×ng / mL), half-life (t1 / 2 = 2.7h) and (Cmax = 5.75μM) in plasma. Analysis of the content of compound 1 in the hippocampus of mice showed a brain-to-plasma ratio of 0.33. CCK8 detection showed that compound 1 and the control antiepileptic drug lamotrigine (LTG) had no toxic effects on the proliferation of HepG2 cells at the tested concentrations (0.078-40μM). The median lethal dose (LD50) of acute toxicity of compound 1 is 596mg / kg.

[0188] Example 7 Targeting specificity verification of compound 1

[0189] To test the targeting specificity of compound 1 to PSPH in cells, we performed single-point mutations in the amino acids that interact with PSPH and compound 1, and these interaction sites showed a high degree of evolutionary conservation ( Fig.13 The results showed that mutations in Asp22 and Lys158 completely abolished the activity of PSPH ( Fig.13 b). The effect of Ala51Val mutation on PSPH activity is negligible. Gly110Ala and Ala51Val / Gly110Ala double mutations can reduce the activity of PSPH by about 40% ( Fig.13b). The results of microcalorimetry experiments showed that the Kd of the Ala51Val / Gly110AlaPSPH mutant (PSPH-MT) was 6.1 μM ( Fig.13 c), indicating that the binding affinity of the Ala51Val / Gly110AlaPSPH mutant to compound 1 was reduced by 26-fold compared with wild-type PSPH. When we knocked out endogenous wild-type PSPH in primary astrocytes and overexpressed PSPH-MT using a viral vector, we found that the inhibitory effect of 1 μM compound 1 on cellular and extracellular L-serine levels was abolished ( Fig.13 This result suggests that compound 1 affects serine production through PSPH.

[0190] In addition, we evaluated the activity of compound 1 against 20 protein tyrosine phosphatases and 5 protein serine / threonine phosphatases. Compared with PSPH, compound 1 had weak or no effects on all tested targets ( Figure 14-15 ). In conclusion, compound 1 downregulated the levels of L-serine and D-serine by inhibiting PSPH.

[0191] Example 8 Effect of Compound 1 in Reducing Spontaneous Seizures in Mice with Temporal Lobe Epilepsy

[0192] A chronic temporal lobe epilepsy mouse model was established by unilateral hippocampal injection of kainic acid (KA) using the stereotaxic method. The detailed parameters were as follows: the hippocampal positioning injection coordinates with Bregma as the origin were: -1.8, -1.8, -2.0 mm, and 12 mg / kg of kainic acid was injected unilaterally. The anti-epileptic effect of compound 1 on mice with spontaneous epileptic seizures was tested: 4 weeks after kainate-induced status epilepticus, mice were implanted with wired EEG electrodes to monitor the mice's daily spontaneous electroclinical seizures (SESs) and convulsive behavior. The EEG recordings in the first three weeks were used to assess the baseline frequency of SESs in each mouse. In the next six weeks, mice were treated with solvent (control group), lamotrigine (LTG), or compound 1 ( Fig.16 As mentioned above, spontaneous electroclinical seizures increased gradually over time in the vehicle control group ( Fig.16 Lamotrigine treatment resulted in an abrupt decrease in SESs in the first week, but the efficacy gradually decreased as treatment continued ( Fig.16 c, f, h in Figure 3). This result is consistent with previous reports that the kainic acid chronic TLE mouse model showed resistance to traditional anti-epileptic drugs. Although 2 mg / kg of compound 1 treatment prevented the increase in SESs, the frequency of SESs was still higher than the baseline ( Fig.16A higher dose of compound 1 (4 mg / kg) significantly suppressed SESs to 0.44 ± 0.15 per day, a mean decrease of 75% compared with the baseline (1.79 ± 1.1). Fig.16 Overall, these data suggest that compound 1 may be a potential therapeutic agent for temporal lobe epilepsy.

Claims

1. Use of PSPH inhibitors in the preparation of drugs for preventing, treating or ameliorating nervous system diseases.

2. The use according to claim 1, characterized in that The PSPH inhibitor prevents, treats or improves nervous system diseases by inhibiting the activity of phosphoserine phosphatase, thereby inhibiting the production and release of L-serine in astrocytes, and then inhibiting the upregulation of D-serine.

3. The use according to claim 1, characterized in that The nervous system disease is selected from at least one of cognitive impairment, intellectual disability, cerebellar insufficiency, epilepsy, neurodevelopmental disorder, dementia, autism spectrum disorder, Down syndrome, Rett syndrome, fragile X syndrome, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and ischemia-reperfusion injury.

4. The use according to claim 3, characterized in that The nervous system disease is selected from at least one of stroke, epilepsy, chronic pain, Parkinson's disease, Alzheimer's disease, Huntington's disease, and ischemia-reperfusion injury.

5. The use according to claim 4, characterized in that The nervous system disease is epilepsy.

6. The use according to claim 5, characterized in that The nervous system disease is temporal lobe epilepsy.

7. The use according to claim 1, characterized in that The PSPH inhibitor is a compound of formula I or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof: In the formula, R1 is selected from substituted or unsubstituted C 1-8 Alkyl, C 3-8 Cycloalkyl, aryl, heterocyclic, halogen, C 1-8 Alkoxy, the substituted substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6-membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6-membered heterocyclic group containing oxygen and / or nitrogen; R2 is -CH=N- or -CH2CH 2- .

8. The use according to claim 7, characterized in that R1 is selected from substituted or unsubstituted C 1-5 Alkyl, C 3-5 Cycloalkyl, aryl, 3-6 membered heterocyclic group, halogen, C 1-5 Alkoxy, the substituted substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6 membered heterocyclic group containing oxygen and / or nitrogen.

9. The use according to claim 8, characterized in that R1 is selected from C 1-5 Alkyl, C 3-5 Cycloalkyl, phenyl, halogen, C 1-5 Alkoxy.

10. The use according to claim 9, characterized in that R1 is selected from C 1-3 Alkyl, phenyl, halogen, C 1-3 Alkoxy.

11. The use according to claim 10, characterized in that R1 is selected from C 1-3 Alkyl, halogen, C 1-3 Alkoxy.

12. The use according to claim 11, characterized in that R1 is selected from halogen, C 1-3 Alkoxy.

13. The use according to claim 12, characterized in that R1 is selected from F, Cl, Br, C 1-3 Alkoxy.

14. The use according to claim 13, characterized in that R1 is selected from F and methoxy, preferably F.

15. The use according to claim 14, characterized in that The compound of formula I is selected from the following compounds:

16. The use according to claim 15, characterized in that The compound of formula I is selected from the following compounds:

17. Use of D-serine inhibitors in the preparation of drugs for treating nervous system diseases. The nervous system disease has the same definition as the disease defined in the use according to any one of claims 3 to 6.

18. The use according to claim 17, characterized in that The D-serine inhibitor comprises a compound of formula I or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof, and the compound of formula I has the same definition as the compound of formula I defined in the use described in any one of claims 7 to 16.

19. Use of L-serine inhibitors in the preparation of drugs for treating nervous system diseases. The nervous system disease has the same definition as the disease defined in the use according to any one of claims 3 to 6.

20. The use according to claim 19, characterized in that The L-serine inhibitor can inhibit the production and release of L-serine in astrocytes.

21. The use according to claim 19, characterized in that The L-serine inhibitor comprises a compound of formula I or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof, and the compound of formula I has the same definition as the compound of formula I defined in the use described in any one of claims 7 to 16.

22. A compound or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, characterized in that: The compound structural formula is as follows: Wherein, R1 has the same definition as described in any one of claims 7-14.

23. The compound according to claim 22 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, characterized in that: The compound structural formula is as follows:

24. A method for preparing the compound according to claim 22 or 23, characterized in that: The following steps are involved:

25. The preparation method according to claim 24, characterized in that: The following steps are involved: (1) Add compound 1, a base, a catalyst and (Boc)2O to a solvent, and react at 20-40° C. for 1-5 hours to obtain compound 2; (2) adding NaBH4 to the solution of compound 2 at a temperature of -10-10°C and reacting for 0.2-1 hour to obtain compound 3; (3) Compound 3 is dissolved in a solvent, cooled to -5-5°C, and a base and MsCl (methylsulfonyl chloride) are added in sequence, and the mixture is reacted at -10-10°C for 2-4 hours to obtain compound 4; (4) Compound 4 is added with a solvent and a reducing agent at -5-5°C under an inert atmosphere, and the mixture is heated to 20-40°C for reaction for 1-3 hours to obtain compound 5; (5) Compound 5 is mixed with potassium cyanate and a solvent, and reacted at 55-75° C. in an inert atmosphere for 2-4 hours to obtain a product.

26. A medicament comprising the compound of claim 22 or 23 or its stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug.

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