A class of carbamate compounds and use thereof in the preparation of neuroprotective agents or cholinesterase inhibitors

Abstract

The present application relates to a kind of urethane compounds with the structure shown in general formula (I), its preparation method and its application in medical field.The compound and its pharmaceutically acceptable salt, solvate, stereoisomer or prodrug, show significant selective butyrylcholinesterase inhibitory activity, and have significant protection effect on glutamate-induced neuron damage.The preparation route of the compound is also disclosed.The pharmacological activity screening result shows that the compound can be used as cholinesterase inhibitor and / or neuroprotective agent, for preparing drug for treating, preventing or relieving neurodegenerative disease or nervous system disease related to abnormal function of cholinergic system and / or neuron damage, including Alzheimer's disease, Lewy body dementia, progressive supranuclear palsy, vascular dementia, myasthenia gravis, glaucoma, cognitive dysfunction or movement disorder caused by central or peripheral nervous system degenerative disease.The compound of the present application can also be used for preparing neuroprotective agent.(I)

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a class of carbamate compounds of general formula (I) or their pharmaceutically acceptable salts, stereoisomers, or mixtures. These compounds possess selective butyrylcholinesterase inhibitory activity and neuroprotective effects, and can be used as cholinesterase inhibitors and / or neuroprotective agents in the preparation of drugs for the treatment, prevention, or alleviation of neurodegenerative diseases such as Alzheimer's disease, Lewy body dementia, and vascular dementia, as well as cognitive or motor impairments caused by damage to the central or peripheral nervous system. Background Technology

[0002] Alzheimer's disease (AD) is a serious neurodegenerative disease that severely impacts the health of older adults. It is the most common type of dementia, accounting for approximately 50%-75% of all dementia cases. Clinical manifestations include progressive cognitive decline, memory loss, and behavioral disturbances. Key pathological features include senile plaques formed by β-amyloid protein deposition, neurofibrillary tangles formed by hyperphosphorylated tau protein aggregation, neuroinflammatory responses, and neuronal loss. With the increasing aging of the global population, the number of AD patients is rapidly rising, and is projected to exceed 100 million by 2050. Currently, the exact pathogenesis of AD remains unclear, and existing treatments can only alleviate symptoms and cannot stop disease progression. Based on the "cholinergic hypothesis," acetylcholinesterase (AChE) inhibitors, such as donepezil and galantamine, improve cognitive function by increasing acetylcholine levels in the synaptic cleft and are currently a commonly used class of symptomatic treatment drugs in clinical practice. However, these drugs are less effective in advanced Alzheimer's disease (AD) patients because, as the disease progresses, AChE activity in the brain gradually decreases, while butyrylcholinesterase (BuChE) activity increases compensatorily. In advanced AD patients, BuChE can become the main enzyme for hydrolyzing acetylcholine in the brain. Therefore, selectively inhibiting BuChE can effectively increase acetylcholine levels while reducing the peripheral side effects that may be caused by non-selective AChE inhibition, making it an important direction for the development of anti-AD drugs.

[0003] Besides cholinergic system dysfunction, glutamate-mediated excitotoxicity is one of the core mechanisms of neuronal damage in Alzheimer's disease (AD) and other neurodegenerative diseases. Excessive glutamate activation of NMDA receptors leads to calcium ion influx, mitochondrial dysfunction, and oxidative stress, ultimately resulting in neuronal apoptosis or necrosis. Notably, glutamate excitotoxicity is also a key pathological link in ischemic stroke (cerebral ischemia-reperfusion injury)—after cerebral ischemia, a large amount of extracellular glutamate accumulates, overstimulating NMDA receptors and triggering delayed neuronal death, a significant cause of post-stroke neurological deficits. Therefore, compounds that can antagonize glutamate damage hold promise not only for neurodegenerative diseases but may also have a protective effect against neurological damage caused by ischemic stroke. Currently, only memantine is used clinically as an NMDA receptor antagonist for moderate to severe AD, but its protection against glutamate damage is incomplete, and it lacks cholinesterase regulatory activity.

[0004] Current AD treatments have significant limitations: AChE inhibitors such as donepezil and galantamine show decreased efficacy in advanced patients and have significant peripheral side effects; while rivastigmine can inhibit both AChE and BuChE, its BuChE inhibition ability is low, and its selectivity for both enzymes is not high, potentially causing cholinergic-related side effects; currently, there are few reported highly selective BuChE inhibitors, and none have entered clinical use. Furthermore, drugs that simultaneously improve cholinergic function and provide glutamate neuroprotection are extremely rare; amimantan acts on the glutamate pathway but lacks cholinesterase inhibitory activity. In addition, for ischemic stroke, current thrombolysis and thrombectomy treatments have a narrow time window and lack effective neuroprotective drugs, while the glutamate injury mechanism provides an important target for the development of novel neuroprotective agents.

[0005] Therefore, developing a class of novel compounds with selective butyrylcholinesterase inhibitory activity and glutamate-induced neuronal damage protection is of significant clinical and scientific value for the preparation of drugs to treat neurodegenerative diseases such as Alzheimer's disease, Lewy body dementia, and vascular dementia, and potentially extend to neurological damage caused by ischemic stroke. Based on this need, this invention provides a class of carbamate compounds represented by general formula (I), their preparation method, and their pharmaceutical uses. Summary of the Invention

[0006] This invention aims to provide a class of novel carbamate compounds represented by general formula (I), their preparation methods, and their applications in the pharmaceutical field. These compounds exhibit selective butyrylcholinesterase inhibitory activity and protective effects against glutamate-induced neuronal damage. They can be used as cholinesterase inhibitors and / or neuroprotective agents for the preparation of drugs to treat, prevent, or alleviate neurodegenerative diseases or nervous system disorders related to cholinergic system dysfunction and / or neuronal damage. These diseases include, but are not limited to, Alzheimer's disease, Lewy body dementia, progressive supranuclear palsy, vascular dementia, myasthenia gravis, glaucoma, and cognitive or motor impairments caused by central or peripheral nervous system degeneration. This invention also discloses the preparation routes of these compounds and pharmaceutical compositions containing these compounds.

[0007] A carbamate compound, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, enantiomer, diastereomer, racemate, or mixture thereof, wherein the carbamate compound, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, enantiomer, diastereomer, racemate, or mixture thereof, is represented by general formula (I): (I) In formula (Ⅰ), X is CH2, O, or S; n is 0 or 1; W1, W2, W3, W4 and W5 are each independently CH or N, and the total number of N does not exceed 2; R1 is a mono- or di-(C1-C4 alkyl) substituted amino group, or an unsubstituted or substituted 5- to 7-membered nitrogen heterocyclic group substituted by 1-2 substituents independently selected from halogen, C1- to C4 alkyl, hydroxyl, hydroxymethyl, trifluoromethyl, carboxyl, C2- to C4 acyl, amide, etc., wherein the nitrogen heterocyclic group contains 0-1 additional heteroatom selected from O, S, N.

[0008] R2 is selected from hydrogen, halogen, C1-C4 straight-chain or branched alkyl, C2-C4 straight-chain or branched alkenyl, C2-C4 straight-chain or branched alkynyl, C1-C4 alkoxy, C1-C4 alkylcarbonyloxy, cyano, nitro, hydroxy, hydroxymethyl, trifluoromethyl, carboxyl, mercapto, methyl mercapto, C1-C4 acyl, amide, aminosulfonyl, C1-C4 alkylsulfonyl, or two adjacent R2 substituents together with the carbon atom to which they are attached form a 5- to 7-membered ring; wherein the number of R2 is 0, 1, 2 or 3.

[0009] As a further improvement to the above scheme, the compound is selected from the group consisting of: As a further improvement to the above scheme, the compound has butyrylcholinesterase selective inhibitory activity, and the drug is used as a cholinesterase inhibitor for the treatment, prevention or relief of Alzheimer's disease or functional impairments related to cholinergic system dysfunction.

[0010] As a further improvement to the above scheme, the compound has selective inhibitory activity against butyrylcholinesterase and / or protective effect against glutamate-induced neuronal damage. The drug is used as a cholinesterase inhibitor and / or neuroprotective agent for the treatment, prevention or relief of neurodegenerative diseases or nervous system diseases associated with cholinergic system dysfunction and / or neuronal damage.

[0011] As a further improvement to the above solution, the following steps are included: (1) In a protic solvent and in the presence of an inorganic base, an amidine compound J is reacted with a β-keto ester compound Y to obtain a pyrimidine compound JY; wherein the protic solvent is selected from one or more of methanol, ethanol, and isopropanol; and the inorganic base is selected from one or more of potassium carbonate, sodium carbonate, cesium carbonate, sodium bicarbonate, sodium hydroxide, potassium hydroxide, potassium phosphate, sodium phosphate, dipotassium hydrogen phosphate, and disodium hydrogen phosphate. (2) In a polar aprotic solvent, in the presence of an organic base, a pyrimidine compound JY is reacted with a carbamoyl chloride compound d to obtain a carbamate compound JY-d; wherein the polar aprotic solvent is selected from one or more of acetonitrile, tetrahydrofuran, dioxane, dichloromethane, chloroform, N,N-dimethylformamide, dimethyl sulfoxide, and N-methylpyrrolidone; wherein the organic base is selected from one or more of triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, and pyridine; In the above reaction formulas, the definitions of R1, R2, X, W1-W4 substituents and n are as described in claim 1.

[0012] As a further improvement to the above scheme, the pharmaceutical composition comprises a therapeutically effective amount of one or more carbamate compounds as described in claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, enantiomer, diastereomer, racemate, or mixture thereof, and optionally a pharmaceutically acceptable carrier.

[0013] As a further improvement to the above scheme, the disease is selected from one or more of Alzheimer's disease, Lewy body dementia, progressive supranuclear palsy, vascular dementia, myasthenia gravis, glaucoma, and cognitive or motor impairment caused by degenerative lesions of the central or peripheral nervous system. Detailed Implementation

[0014] Within the scope of this invention, the above-mentioned technical features and the technical features specifically described in the embodiments below can be arbitrarily combined to form new or preferred technical solutions, and these combinations are all within the scope of this invention.

[0015] Method of drug administration: Since the carbamate compounds represented by general formula (I) of this invention have selective butyrylcholinesterase inhibitory activity and / or protective effects against glutamate-induced neuronal damage, these compounds, as well as pharmaceutically acceptable salts, solvates, stereoisomers or mixtures thereof, and pharmaceutical compositions containing these compounds as the main active ingredient, can be used to treat, prevent or alleviate neurodegenerative diseases or nervous system diseases associated with cholinergic system dysfunction and / or neuronal damage.

[0016] The pharmaceutical compositions of the present invention comprise, within a safe and effective range, the compound of the present invention or a pharmaceutically acceptable salt, solvate, stereoisomer, or mixture thereof, and a pharmaceutically acceptable excipient or carrier. Herein, "safe and effective range" means that the amount of the compound is sufficient to significantly improve the condition or exert the intended pharmacological effect without causing serious side effects. Typically, for an adult weighing 60 kg, the daily dose of the compound of the present invention is 1 mg to 2000 mg, and the specific dose may be adjusted by a physician based on factors such as route of administration, patient age, weight, and severity of condition.

[0017] The "pharmaceutically acceptable salt" described in this invention refers to a conventional non-toxic salt formed by the reaction of a compound of general formula (I) with an inorganic or organic acid. The inorganic acid includes one or more of hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, aminosulfonic acid, and phosphoric acid; the organic acid includes one or more of citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, maleic acid, malic acid, malonic acid, fumaric acid, succinic acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pyric acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, or hydroxyethanesulfonic acid.

[0018] The term "pharmaceutically acceptable carrier" as used in this invention refers to one or more compatible solid or semi-solid, liquid filler or gel material that is suitable for human use and has sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with and with the compounds of this invention without significantly reducing the efficacy of the compounds.

[0019] Suitable pharmaceutical carriers include, but are not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), phosphates, glycerol, sorbic acid, potassium sorbate, vegetable oils, mixtures of glycerides of saturated vegetable fatty acids, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, calcium sulfate, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, beeswax, lanolin, gelatin, talc, solid lubricants, emulsifiers, wetting agents, colorants, flavoring agents, stabilizers, antioxidants, preservatives, and pyrogen-free water.

[0020] There are no particular limitations on the administration of the compound or pharmaceutical composition of formula (I) of the present invention. Representative administration methods include oral, intramuscular, intravenous, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, transdermal, conjunctival, local, or implant-based administration.

[0021] Solid dosage forms for oral administration include capsules, tablets (including uncoated and coated tablets), pills, powders, and granules. In these solid dosage forms, the active compound of formula (I) is mixed with at least one conventional inert excipient or carrier, said excipient or carrier comprising: (a) a filler or compatibilizer, such as starch, lactose, sucrose, glucose, mannitol, or silica; (b) a binder, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, or gum arabic; (c) a humectant, such as glycerin; (d) a disintegrant, such as agar, calcium carbonate, potato starch or cassava starch, alginate, complex silicates, or sodium carbonate; (e) a slow solvent, such as paraffin wax; (f) an absorption accelerator, such as a quaternary ammonium salt compound; (g) a wetting agent, such as cetyl alcohol or glyceryl monostearate; (h) an adsorbent, such as kaolin; and (i) a lubricant, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof.

[0022] Buffers may also be included in capsules, tablets, and pills. The aforementioned solid dosage forms may be prepared using coatings and shell materials, such as enteric coatings or other coating materials known in the art. Coatings can be used to control the release of the active ingredient at specific sites in the digestive tract. Examples of encapsulating components that may be used include polymeric substances and waxes. Where necessary, the active compound may be formed into microcapsules with one or more of the aforementioned excipients.

[0023] Liquid dosage forms for oral administration include pharmaceutically acceptable solutions, emulsions, suspensions, syrups, or elixirs. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, dimethyl sulfoxide, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil, or mixtures of these substances.

[0024] In addition to inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, and aroma agents. The suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol, dehydrated sorbitan esters, microcrystalline cellulose, aluminum methacrylate, agar, or mixtures of these substances.

[0025] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or aqueous solutions, dispersions, suspensions, emulsions, lyophilized products, and sterile powders, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions, and suitable aqueous or non-aqueous carriers and diluents.

[0026] Dosage forms suitable for other routes of administration include: nasal drops / solutions, sprays; tablets or capsules for administration to the tongue, sublingual or buccal; suppositories; preparations for use in the eyes and ears; vaginal capsules; aqueous suspensions (lotions, shaken mixtures); lipophilic suspensions; ointments; creams; emulsions; pastes; powders or implants.

[0027] The active ingredient of formula (I) can be converted into the above-described delivery form using methods known in the art, which can be achieved by using inert, non-toxic, and suitable pharmaceutical excipients, including: carriers (such as microcrystalline cellulose), solvents (such as liquid polyethylene glycol), emulsifiers (such as sodium dodecyl sulfate), dispersants (such as polyvinylpyrrolidone), synthetic and natural biopolymers, stabilizers (such as antioxidants), colorants (such as inorganic pigments), flavoring agents, and / or taste masking agents. Where suitable, the active ingredient may be present in one or more of the above-described carriers in the form of microencapsulation.

[0028] In addition to the compounds of formula (I) of the present invention, the above-mentioned pharmaceutical preparations may also contain one or more other active pharmaceutical ingredients that have neuroprotective, cholinesterase inhibitory or symptomatic therapeutic effects.

[0029] When using the pharmaceutical compositions of the present invention, a safe and effective amount of the compound of the present invention is administered to the mammal (preferably a human) requiring treatment. For an adult weighing 60 kg, the daily dose is typically 1 mg to 2000 mg. The specific dose should be determined by the clinician based on factors such as route of administration, patient age, weight, severity of condition, and individual differences. It is generally recommended to start with a lower dose and gradually adjust to the optimal therapeutic dose.

[0030] The main advantages of this invention include: providing a class of novel carbamate compounds represented by general formula (I) that have selective butyrylcholinesterase inhibitory activity and protective effect against glutamate-induced neuronal damage, and can be used as cholinesterase inhibitors and / or neuroprotective agents for the preparation of drugs for the treatment, prevention or relief of neurodegenerative diseases or nervous system diseases related to cholinergic system dysfunction and / or neuronal damage.

[0031] It should be understood that after reading the above description of this invention, those skilled in the art can make various alterations or modifications to this invention, but these equivalent forms also fall within the protection scope defined by the appended claims.

[0032] English abbreviations and their full Chinese names Through extensive and in-depth research and screening, the inventors have developed a class of active ingredients that can effectively inhibit cholinesterase, namely compounds represented by general formula (I) or their pharmaceutically acceptable salts, solvates, hydrates, enantiomers, diastereomers, or racemates. Experiments have shown that the active ingredients of this invention can efficiently inhibit hAChE activity, thus making them suitable for treating diseases related to nervous system disorders. Based on this, the present invention was completed.

[0033] The technical solution of the present invention will be clearly and completely described below. However, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0034] the term Unless otherwise stated, the following terms used in this specification have the following meanings: The term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0035] The term "C1-C4 straight-chain or branched alkyl" refers to a saturated straight-chain or branched hydrocarbon group having 1 to 4 carbon atoms, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0036] The term "C2-C4 straight-chain or branched alkenyl" refers to a straight-chain or branched hydrocarbon group having 2 to 4 carbon atoms and containing at least one carbon-carbon double bond, including but not limited to vinyl, propenyl, and butenyl.

[0037] The term "C2-C4 straight-chain or branched alkenyl" refers to a straight-chain or branched hydrocarbon group having 2 to 4 carbon atoms and containing at least one carbon-carbon double bond, including but not limited to vinyl, propenyl, and butenyl.

[0038] The term "C1-C4 alkoxy" refers to straight-chain or branched alkoxy groups having 1 to 4 carbon atoms, i.e., -O-(C1-C4 alkyl), including but not limited to methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy.

[0039] The term "C1-C4 alkyl carbonyloxy" refers to an alkyl carbonyloxy group having 1 to 4 carbon atoms, i.e., -OC(=O)-(C1-C4 alkyl), including but not limited to acetoxy (-OCOCH3), propionyloxy, and butyryloxy.

[0040] The term "C1-C4 acyl" refers to an alkyl carbonyl group having 1 to 4 carbon atoms, i.e., -C(=O)-(C1-C4 alkyl), including but not limited to formyl, acetyl, propionyl, and butyryl.

[0041] The term "amide group" refers to -C(=O)-NH2, or -C(=O)-NH (C1-C4 alkyl) or -C(=O)-N (C1-C4 alkyl)2, in which one or two hydrogens are replaced by C1-C4 alkyl groups.

[0042] The term "aminosulfonyl" refers to -SO2-NH2.

[0043] The term "C1-C4 alkylsulfonyl" refers to an alkylsulfonyl group having 1 to 4 carbon atoms, i.e., -SO2-(C1-C4 alkyl), including but not limited to methanesulfonyl (-SO2CH3), ethanesulfonyl, and propanesulfonyl.

[0044] The term "mono- or di-(C1-C4 alkyl) substituted amino" refers to an amino group (-NH2) in which one or two hydrogen atoms are substituted by a C1-C4 alkyl group to form -NH(C1-C4 alkyl) or -N(C1-C4 alkyl)2, including but not limited to methylamino, ethylamino, dimethylamino, diethylamino, and ethylmethylamino.

[0045] The term "5- to 7-membered nitrogen heterocyclic group" refers to a saturated or partially unsaturated 5-, 6-, or 7-membered monocyclic group containing at least one nitrogen atom on the ring and may contain 0 to 1 additional heteroatoms selected from O, S, and N. One or more hydrogen atoms on the ring carbon atom may be replaced by substituents selected from halogens, C1-C4 alkyl groups, hydroxyl groups, hydroxymethyl groups, trifluoromethyl groups, carboxyl groups, C2-C4 acyl groups, and amide groups. Exemplary unsubstituted nitrogen heterocyclic groups include, but are not limited to, pyrrolidinyl, piperidinyl, piperazineyl, morpholinyl, heptyl, imidazoalkyl, and pyrazolyl groups.

[0046] The term "pharmaceutically acceptable salt" refers to a salt formed by the compound of this invention with a pharmaceutically acceptable inorganic or organic acid, or a salt formed with a pharmaceutically acceptable base. The inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; the organic acids include formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, benzoic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc. The bases include sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium carbonate, sodium bicarbonate, ammonia, organic amines (such as triethylamine, methylamine, ethylamine, ethanolamine), etc.

[0047] The term "solvent" refers to a substance formed by the combination of the compound of this invention with stoichiometric or non-stoichiometric solvent molecules through intermolecular forces; when the solvent is water, it is called a "hydrate".

[0048] The term "stereoisomer" refers to isomers with different spatial configurations due to chiral centers, double bonds, or cyclic structures in a molecule, including enantiomers, diastereomers, cis-trans isomers, etc.

[0049] The term "racemate" refers to a mixture of equal amounts of a pair of enantiomers, which is usually optically inactive.

[0050] The term "pharmaceutical composition" refers to a composition containing the compound of the present invention or its pharmaceutically acceptable salt, solvate, stereoisomer, etc., and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0051] The term "therapeutic effective amount" refers to an amount sufficient to produce the desired biological or medical response in a subject, such as an amount sufficient to improve symptoms, slow disease progression, or prevent disease onset.

[0052] Example 1: Synthesis of compound JY029 (2-(4-methoxyphenyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-4-yldimethylcarbamate) 4-Methoxybenzomididine (J4, 0.54 g, 3 mmol), methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (Y1, 0.50 g, 3 mmol), and 15 ml of anhydrous ethanol were added to a 50 ml round-bottom flask. Anhydrous potassium carbonate (0.83 g, 6 mmol) was then added. The mixture was heated at 85 °C for 20 h. The entire reaction solution was transferred, 2 g of silica gel was added, and the mixture was stirred until dry. Column chromatography (DCM:MeOH = 15:1) was performed. The resulting chromatogram was concentrated under reduced pressure and dried under vacuum to obtain a white solid JY028 (0.50 g, 61%). ¹H NMR (400 MHz, DMSO) δ 12.34–12.85 (s, 1H), 8.08 (d, J = 8.4 Hz, 2H), 7.05 (d, J = 8.4 Hz, 2H), 3.83 (s, 1H). 3H), 3.51 (s, 3H), 2.96 – 2.80 (m, 4H).13C NMR (101 MHz, DMSO) δ 161.03,158.75, 155.58, 129.03, 128.92, 115.37, 113.49, 113.13, 55.25, 32.70, 24.74,23.27. HRMS (ESI+) m / z calcd for C14H15N2O2S+ 275.0854 found 275.0841 Da. JY028 (0.137 g, 0.5 mmol), 10 ml of 1,4-dioxane, and DIPEA (0.33 g, 2.55 mmol) were added to a 50 mL round-bottom flask. Then, dimethylcarbamoyl chloride (d7, 0.11 g, 1 mmol) was added. The mixture was heated to 65 °C for 18 h. The entire reaction solution was transferred, and 2 g of silica gel was added and stirred until dry. The solution was then subjected to column chromatography (DCM:MeOH = 50:1). The resulting chromatogram was concentrated under reduced pressure to obtain a colorless oil. The oil was dissolved in 10 ml of methanol and sonicated, resulting in a white precipitate. The precipitate was concentrated again under reduced pressure and dried under vacuum to obtain a white solid, JY029 (0.14 g, 81%). ¹H NMR (400 MHz, DMSO) δ 8.23 ​​(d, J = 8.8 Hz, 2H), 7.05 (d, J = 8.9 Hz, 2H). 13C NMR (101 MHz, DMSO) δ HRMS (ESI+) m / z calcd for C17H20N3O3S+ 346.1225 found 346.1213 Da. Example 2: Synthesis of compound JY032 (2-(4-methoxyphenyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-4-ylpiperidine-1-carboxylate) 2-(4-methoxyphenyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-4-ol (JY028, 0.137 g, 0.5 mmol), 1,4-dioxane (10 ml), and DIPEA (0.33 g, 2.55 mmol) were added to a 50 mL round-bottom flask. Piperidine-1-formyl chloride (d21, 0.148 g, 1 mmol) was then added. The mixture was heated to 65 °C for 18 h. The entire reaction mixture was transferred, and 2 g of silica gel was added and stirred until dry. The mixture was then subjected to column chromatography (DCM:MeOH = 40:1). The resulting chromatogram was concentrated under reduced pressure to obtain a pale yellow oil. The oil was dissolved in 10 ml of methanol and sonicated, resulting in a white precipitate. The precipitate was concentrated again under reduced pressure and dried under vacuum to obtain a white solid JY032 (0.16 g, 83%). ¹H NMR (400 MHz, CDCl3) δ 8.39 (d, J = 8.4 Hz,2H), 6.96 (d, J = 8.4 Hz, 2H), 3.87 (s, 3H), 3.73 (s, 2H), 3.68 (s, 1H), 3.58(s, 4H), 3.40 (s, 1H), 3.35 (s, 2H), 2.99 (t, J = 5.1 Hz, 2H), 1.54 (d, J =13.2 Hz, 2H). 13C NMR (101 MHz, CDCl3) δ 161.81, 129.97, 129.95, 129.82,129.59, 116.11, 113.73, 55.82, 55.75, 55.37, 33.77, 30.87, 30.72, 29.95,29.34, 29.30, 25.58, 23.46. HRMS (ESI+) m / z calcd for C20H24N3O3S + 386.1538found 386.1527 Da. Example 3: Synthesis of compound JY039 (2-(3-bromophenyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-4-yldimethylcarbamate) 3-Bromobenzamide (J6, 0.68 g, 3 mmol), methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (Y1, 0.50 g, 3 mmol), and 15 ml of anhydrous ethanol were added to a 50 ml round-bottom flask. Anhydrous potassium carbonate (0.83 g, 6 mmol) was then added, and the mixture was heated at 85 °C for 20 h. The entire reaction mixture was transferred, 2 g of silica gel was added, and the mixture was stirred until dry. Column chromatography (DCM:MeOH = 18:1) was performed. The resulting chromatogram was concentrated under reduced pressure and dried under vacuum to obtain a white solid JY038 (0.52 g, 54%). ¹H NMR (400 MHz, DMSO) δ 12.71–12.93 (m, 1H), 8.27 (s, 1H), 8.08 (d, J = 7.7 Hz, 1H), 7.77 (d, J = 8.0 Hz, 1H). 7.48 (t, J = 7.9 Hz, 1H), 3.53 (s, 2H), 2.89 (s, 4H). 13C NMR (101MHz, DMSO) δ 134.21, 134.06, 130.90, 130.77, 130.32, 130.14, 126.56, 126.00,125.40, 121.88, 32.70, 24.41, 22.67. HRMS (ESI+) m / z calcd for C13H12BrN2OS +322.9854 found 322.9851 Da. Add JY038 (0.162 g, 0.5 mmol), 10 mL of 1,4-dioxane, and TEA (0.26 g, 2.55 mmol) to a 50 mL round-bottom flask, then add dimethylcarbamoyl chloride (d7, 0.11 g, 1 mmol). Heat the mixture at 65 °C for 14 h. Transfer the entire reaction solution, add 2 g of silica gel, stir until dry, and perform column chromatography (DCM:MeOH = 50:1). Concentrate the resulting chromatogram under reduced pressure to obtain a colorless oily substance. Add 0.5 mL of 4M dioxane hydrochloride, sonicate, and a white precipitate forms. Concentrate again under reduced pressure, and dry under vacuum to obtain a white solid JY039, which is the hydrochloride salt (0.16 g, 68%). ¹H NMR (400 MHz, DMSO) δ 8.39 (d, J = 1.6 Hz, 1H), 8.27 (d, J = 7.9 Hz, 1H), 7.75 (d, J = 7.9 Hz, 1H), 7.50 (t, J = 7.9 Hz, 1H), 3.75 (d, J = 11.7 Hz, 2H), 3.19 (dd, J = 11.5, 5.6Hz, 2H), 3.08 (s, 3H), 3.03 (t, J = 5.9 Hz, 2H), 2.98 (s, 3H). 13C NMR (101MHz, CDCl3) δ 167.75, 162.97, 160.78, 152.27, 138.83, 133.61, 131.23, 129.98,126.91, 122.71, 117.55, 36.94, 33.75, 25.45, 23.54. HRMS (ESI+) m / z calcd forC16H17BrN3O2S + 396.0204 found 396.0190 Da. Example 4: Synthesis of compound JY040 (2-(3-bromophenyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-4-ylethyl(methyl)carbamate) JY038 (0.162 g, 0.5 mmol), 10 mL of 1,4-dioxane, and TEA (0.26 g, 2.55 mmol) were added to a 50 mL round-bottom flask. Ethyl (methyl)carbamoyl chloride (d16, 0.121 g, 1 mmol) was then added, and the mixture was heated at 65 °C for 14 h. The entire reaction mixture was transferred, 2 g of silica gel was added, and the mixture was stirred until dry. Column chromatography (DCM:MeOH = 50:1) was performed. The resulting chromatogram was concentrated under reduced pressure to obtain a colorless oily substance. 0.5 mL of 4M dioxane hydrochloride was added, and the mixture was sonicated. A white precipitate formed. The precipitate was concentrated again under reduced pressure and dried under vacuum to obtain a white solid, JY040, as hydrochloride (0.13 g, 60%). ¹H NMR (400 MHz, DMSO) δ 8.39 (d, J = 1.7 Hz, 1H), 8.27 (d, J = 7.9 Hz). Hz, 1H), 7.75 (d, J = 8.0 Hz,1H), 7.50 (t, J = 7.9 Hz, 1H), 3.74 (d, J = 9.5 Hz, 2H), 3.45 (d, J = 7.1 Hz,1H), 3.38 (d, J = 7.1 Hz, 1H), 3.20 (t, J = 5.7 Hz, 2H), 3.09 – 3.00 (m, 3H), 2.97 (s, 1H), 1.21 (ddd, J = 20.0, 13.0, 7.1 Hz, 3H). 13C NMR (101 MHz, DMSO)δ 168.01, 162.57, 158.85, 150.87, 138.24, 133.59, 130.98, 129.89, 126.46,121.98, 118.05, 43.93, 43.72, 33.94, 33.89, 33.20, 24.34, 22.43, 13.07,12.00. HRMS (ESI+) m / z calcd for C17H19BrN3O2S + 408.0381 found 408.0361 Da. Example 5: Synthesis of compound JY043 (2-(4-bromophenyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-4-yldimethylcarbamate) In a 50 ml round-bottom flask, 4-bromobenzamide (J7, 0.68 g, 3 mmol), methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (Y1, 0.50 g, 3 mmol), and 15 ml of 1,4-dioxane were added. Anhydrous potassium carbonate (0.83 g, 6 mmol) was then added. The mixture was heated to 80 °C for 13 h. The entire reaction solution was transferred, and 2 g of silica gel was added and stirred until dry. The solution was then subjected to column chromatography (DCM:MeOH = 15:1). The resulting chromatogram was concentrated under reduced pressure and dried under vacuum to obtain a white solid, JY042 (0.53 g, 55%). ¹H NMR (400 MHz, DMSO) δ 12.73–12.87 (m, 1H), 8.03 (d, J = 8.6 Hz, 2H), 7.72 (d, J = 8.6 Hz, 2H), 3.52 (s, 2H), 2.89 (d, J = 3.8 Hz, 4H). 13C NMR (101 MHz, DMSO) δ 175.33, 158.19,140.28, 131.63, 129.59, 127.06, 118.16, 111.00, 51.52, 24.43, 22.63. HRMS(ESI+) m / z calcd for C13H12BrN2OS+ 322.9854 found 322.9840 Da. Add JY042 (0.162 g, 0.5 mmol), 10 mL of 1,4-dioxane, and NMM (0.27 g, 2.67 mmol) to a 50 mL round-bottom flask, then add dimethylcarbamoyl chloride (d7, 0.11 g, 1 mmol). Heat the mixture at 65 °C for 14 h. Transfer the entire reaction solution, add 2 g of silica gel, stir until dry, and perform column chromatography (DCM:MeOH = 50:1). Concentrate the resulting chromatogram under reduced pressure to obtain a colorless oily substance. Add 0.5 mL of 4M dioxane hydrochloride, sonicate, and a white precipitate forms. Concentrate again under reduced pressure, and dry under vacuum to obtain a white solid JY043, which is the hydrochloride salt (0.15 g, 73%). 1H NMR (400 MHz, DMSO) δ 8.21 (d, J = 8.6 Hz, 2H), 7.73 (d, J = 8.6 Hz, 2H), δ 7.73 (d, J = 8.6 Hz, 2H). 2H), 3.75 (s, 2H), 3.34 (s, 2H), 3.07(s, 3H), 3.02 (t, J = 6.0 Hz, 2H), 2.98 (s, 3H). 13C NMR (101 MHz, DMSO) δ167.99, 162.57, 159.69, 151.49, 135.25, 131.78, 129.62, 124.84, 117.80,36.41, 33.28, 24.46, 22.57. HRMS (ESI+) m / z calcd for C16H17BrN3O2S +394.0225 found 394.0209 Da. Example 6: Synthesis of compound JY052 (2-(p-tolyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-4-ylethyl(methyl)carbamate) 4-Methylbenzamide (J9, 0.44 g, 2.6 mmol), methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (Y1, 0.45 g, 2.6 mmol), and 15 ml of anhydrous ethanol were added to a 50 ml round-bottom flask. Anhydrous potassium carbonate (0.83 g, 6 mmol) was then added. The mixture was heated at 85 °C for 26 h. The entire reaction solution was transferred, and 2 g of silica gel was added and stirred until dry. The solution was then subjected to column chromatography (DCM:MeOH = 15:1). The resulting chromatogram was concentrated under reduced pressure and dried under vacuum to obtain a white solid JY050 (0.55 g, 82%). ¹H NMR (400 MHz, DMSO) δ 12.61–12.73 (m, 1H), 7.99 (d, J = 8.0 Hz, 2H), 7.32 (d, J = 8.1 Hz, 2H), 3.52 (s, 2H), 2.95 – 2.80 (m, 4H), 2.37 (s, 3H). 13C NMR (101 MHz, DMSO) δ161.77, 159.64, 151.44, 141.59, 135.71, 129.18, 127.47, 120.81, 32.77, 24.46,22.66, 20.95. HRMS (ESI+) m / z calcd for C14H15N2OS + 259.0905 found 259.0892Da. Add JY050 (0.129 g, 0.5 mmol), 10 mL of 1,4-dioxane, and DIPEA (0.33 g, 2.55 mmol) to a 50 mL round-bottom flask, then add ethyl (methyl)carbamoyl chloride (d16, 0.121 g, 1 mmol). Heat the mixture at 65 °C for 18 h. Transfer the entire reaction solution, add 2 g of silica gel, stir until dry, and perform column chromatography (DCM:MeOH = 50:1). Concentrate the resulting chromatogram under reduced pressure to obtain a yellow oily substance. Add 0.3 mL of 4M dioxane hydrochloride; a white precipitate forms. Concentrate again under reduced pressure and dry under vacuum to obtain a white solid JY052, which is the hydrochloride salt (0.13 g, 71%). ¹H NMR (400 MHz, DMSO) δ 8.17 (d, J = 8.2 Hz, 2H), 7.32 (d, J = 8.1 Hz, 2H), δ 7.32 (d, J = 8.1 Hz, 2H). 2H), 3.73 (s, 2H), 3.45 (dd, J =14.6, 7.4 Hz, 2H), 3.37 (q, J = 7.1 Hz, 1H), 3.17 (t, J = 5.9 Hz, 2H), 3.02(dd, J = 11.1, 5.2 Hz, 3H), 2.96 (s, 1H), 2.37 (s, 3H), 1.26 – 1.13 (m, 4H).13C NMR (101 MHz, DMSO) δ 167.83, 162.58, 160.69, 151.23, 140.85, 133.67,129.29, 127.62, 116.87, 43.74, 33.93, 33.31, 24.51, 22.51, 21.00, 13.15. HRMS(ESI+) m / z calcd for C18H22N3O2S + 344.1433 found 344.1413 Da. Example 7: Synthesis of compound JY055 (2-(p-tolyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-4-ylpiperidine-1-carboxylate) JY050 (0.129 g, 0.5 mmol), 10 mL of 1,4-dioxane, and DIPEA (0.33 g, 2.55 mmol) were added to a 50 mL round-bottom flask. Piperidine-1-formyl chloride (d21, 0.148 g, 1 mmol) was then added, and the mixture was heated at 65 °C for 18 h. The entire reaction mixture was transferred, 2 g of silica gel was added, and the mixture was stirred until dry. Column chromatography (DCM:MeOH = 60:1) was performed. The resulting chromatogram was concentrated to 5 mL under reduced pressure. 0.3 mL of 4M dioxane hydrochloride was added, and the solvent was evaporated to dryness. After vacuum drying, a white solid JY055 was obtained, which was the hydrochloride salt (0.16 g, 82%). ¹H NMR (400 MHz, CDCl₃) δ 8.25 (d, J = 7.6 Hz, 2H), 7.23 (s, 2H), 3.74 (s, 2H). 3.58 (s, 4H), 3.25 (s, 2H), 3.13 (s, 1H), 3.00 (s, 2H), 2.40 (s, 3H), 1.60 (s, 5H). 13C NMR (101 MHz, CDCl3) δ 151.18, HRMS (ESI+) m / z calcd for C20H24N3O2S+ 370.1589 found370.1582 Da. Example 8: Synthesis of compound JY061 (pyrrolidine-1-carboxylic acid 2-(4-fluorophenyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidine-4-yl ester) 4-Fluorobenzoic acid (J10, 0.28 g, 1.6 mmol), methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (Y1, 0.28 g, 1.6 mmol), and 15 ml of anhydrous ethanol were added to a 50 ml round-bottom flask. Anhydrous potassium carbonate (0.83 g, 6 mmol) was then added. The mixture was heated at 85 °C for 20 h. The entire reaction mixture was transferred, and 2 g of silica gel was added and stirred until dry. The mixture was then subjected to column chromatography (DCM:MeOH = 15:1). The resulting chromatogram was concentrated under reduced pressure and dried under vacuum to obtain a white solid JY057 (0.37 g, 88%). ¹H NMR (400 MHz, DMSO) δ 12.73–12.88 (m, 1H), 8.21–8.08 (m, 2H), 7.37 (t, J = 8.8 Hz, 2H), 3.52 (s, 2H), 2.96 – 2.82 (m, 4H). 13C NMR (101 MHz, DMSO) δ 165.26, 161.98,152.50, 137.39, 130.21, 130.13, 128.19, 115.76, 115.54, 32.72, 24.42, 22.64.HRMS (ESI+) m / z calcd for C13H12FN2OS+ 263.0654 found 263.0637 Da. Add JY057 (0.131 g, 0.5 mmol), 10 ml of 1,4-dioxane, and DIPEA (0.33 g, 2.55 mmol) to a 50 mL round-bottom flask, then add pyrrolidine-1-formyl chloride (d19, 0.134 g, 1 mmol). Heat the mixture at 65 °C for 18 h. Transfer the entire reaction solution, add 2 g of silica gel, stir until dry, and perform column chromatography (DCM:MeOH = 60:1). Concentrate the resulting chromatogram under reduced pressure to obtain a yellow oily substance. Dissolve the yellow oily substance in 10 ml of methanol and sonicate. A white precipitate forms. Concentrate again under reduced pressure and dry under vacuum to obtain a white solid JY061 (0.14 g, 78%). ¹H NMR (400 MHz, CDCl₃) δ 8.37 (s, 2H), 7.11 (t, J = 8.0 Hz, 2H), 3.75 (s, 2H), 3.66 – 3.51 (m, 4H), 3.28 (d, J = 26.8 Hz, 5H), 2.99 (s, 2H), 0.87 (s, 1H). 13C NMR (101 MHz, CDCl3) δ 161.28, 150.41, 130.50, 130.41, 115.47, 115.26, 46.81, 44.99, 33.76,25.73, 25.48, 25.00, 24.38, 23.53. HRMS (ESI+) m / z calcd for C18H19FN3O2S +360.1182 found 360.1172 Da. Example 9: Synthesis of compound JY063 (2-(4-fluorophenyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-4-yl diisopropylcarbamate) JY057 (0.131 g, 0.5 mmol), 10 ml of 1,4-dioxane, and DIPEA (0.33 g, 2.55 mmol) were added to a 50 mL round-bottom flask. Then, diisopropylcarbamoyl chloride (d33, 0.164 g, 1 mmol) was added, and the mixture was heated at 65 °C for 15 h. The entire reaction mixture was transferred, 2 g of silica gel was added, and the mixture was stirred until dry. Column chromatography (DCM:MeOH = 50:1) was performed. The resulting chromatogram was concentrated under reduced pressure and dried under vacuum to obtain a white solid, JY063 (0.12 g, 62%). ¹H NMR (400 MHz, CDCl₃) δ 8.43–8.33 (m, 2H), 7.11 (t, J = 8.6 Hz, 2H), 3.75 (s, 2H), 3.24 (t, J = 5.9 Hz, 2H). 3.00 (t, J = 5.9 Hz, 2H), 1.42 – 1.30 (m, 12H). 13C NMR (101 MHz, CDCl3)δ 167.24, 163.28, 161.19, 150.89, 133.10, 130.43, 130.34, 116.52, 115.43,115.21, 33.75, 25.46, 23.57, 21.56, 20.34. HRMS (ESI+) m / z calcd forC20H25FN3O2S+ 390.1652 found 390.1639 Da. Example 10: Synthesis of compound JY115 (2-(3,4-difluorophenyl)-7,8-dihydro-5H-thiopyrano[4,3-d]pyrimidin-4-ylpyrrolidine-1-carboxylate) In a 50 mL round-bottom flask, 3,4-difluorobenzomididine (J18, 0.58 g, 3 mmol), methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (Y1, 0.52 g, 3 mmol), and 15 mL of anhydrous ethanol were added. Then, anhydrous potassium carbonate (0.83 g, 6 mmol) was added. The mixture was heated at 85 °C for 23 h. The entire reaction solution was transferred, and 2 g of silica gel was added and stirred until dry. The solution was then subjected to column chromatography (DCM:MeOH = 15:1). The resulting chromatogram was concentrated under reduced pressure until the solvent was almost completely evaporated. After vacuum drying, a white solid JY112 (0.60 g, 71%) was obtained. ¹H NMR (400 MHz, DMSO) δ 12.62–12.98 (m, 1H), 8.20–8.08 (m, 1H), 7.98 (s, 1H), 7.61 (dd, J = 18.7, 9.9 Hz, 1H), 3.53 (s, 2H), 2.89 (s, 4H). 13C NMR (101MHz, DMSO) δ 150.57, 148.00, 125.08, 125.03, 118.02, 117.84, 116.94, 116.75,32.69, 24.39, 22.64, 20.45, 19.64. HRMS (ESI+) m / z calcd for C13H11F2N2OS+281.0560 found 281.0544 Da. JY112 (0.140 g, 0.5 mmol), 10 mL of 1,4-dioxane, and DIPEA (0.33 g, 2.55 mmol) were added to a 50 mL round-bottom flask. Then, pyrrolidine-1-formyl chloride (d19, 0.134 g, 1 mmol) was added. The mixture was heated at 80 °C for 23 h. The entire reaction solution was transferred, and 2 g of silica gel was added and stirred until dry. The solution was then subjected to column chromatography (DCM:MeOH = 50:1). The resulting chromatogram was concentrated under reduced pressure and dried under vacuum to obtain a white solid, JY115 (0.16 g, 85%). ¹H NMR (400 MHz, CDCl₃) δ 8.25–8.18 (m, 1H), 8.15 (s, 1H), 7.21 (d, J = 8.9 Hz, 1H), 3.75 (s, 2H), 3.65 –3.51 (m, 4H), 3.24 (s, 2H), 3.00 (s, 2H), 2.00 (s, 4H). 13C NMR (101 MHz, CDCl3) δ 171.03, 165.46, 157.18, 155.70, 152.51, 149.87, 126.38, 125.69,120.76, 117.56, 110.96, 49.59, 33.08, 26.11, 25.19, 23.01. HRMS (ESI+) m / zcalcd for C18H18F2N3O2S+ 378.1088 found 378.1075 Da. Following a similar method to Examples 1-10, the following compounds have been synthesized: Example 11: Test of the inhibitory activity of the compound against AChE / BuChE: The micro-DTNB method was used to test the inhibitory activity of carbamate compounds of general formula (Ⅰ) against AChE / BuChE and to calculate the IC50 value of highly active compounds. ATCh is an analog of ACh and can be catalytically hydrolyzed by AChE / BuChE to produce acetic acid and thiocholine iodide (TCh) (see reaction A). TCh can react rapidly and quantitatively with DTNB to produce a yellow anion 5-thio-2-nitrobenzoic acid (RS-) (reaction B). The latter has maximum absorption at a wavelength of 412 nm. The activity of AChE can be calculated by measuring the absorbance (OD value) of the generated RS- to quantify the generated TCh.

[0053] The following is the reaction principle of the micro-DTNB method: This experiment first involved co-incubating AChE / BuChE with a dual concentration of drug (a 1*10-2M drug stock solution was prepared using DMSO and then diluted to the corresponding concentration with PBS), followed by the addition of ATCh for reaction, and finally the addition of DTNB for color development. The enzyme activity rate was calculated by comparing the results with those of normal AChE testing (Table 1). Then, compounds with strong enzyme inhibition ability were selected, and the IC50 value was calculated by measuring the enzyme inhibition rate at multiple concentrations. The specific operating steps are as follows: ① Dilute the AChE / BuChE stock solution 500-2000 times with PBS (0.1M, pH = 7.4, 0.1% BSA) (preliminary experiments confirmed that the OD value measured within 30 min of the reaction between AChE / BuChE and ATCh (3 mM) is linear, and the endpoint OD value generally does not exceed 1.5), and store at 0-4℃. ② Add 20 μL of the diluted enzyme to 10 μL of the corresponding drug concentration (operate in a 96-well plate; the final drug concentration is shown in Table 1), and incubate at 25 ℃ for 10 min. The normal enzyme control is 10 μL of PBS buffer instead of the corresponding drug concentration, and the blank control is 20 μL of PBS buffer instead of the corresponding enzyme (normal and blank controls are set up for each plate). ③ Add 20 μL of ATCh (3 mM) to the enzyme and react at 37 ℃ for 15-20 min. ④ Add 100 μL of DTNB (0.75 mM) and measure the OD value at 412 nM using a microplate reader. Enzyme activity rate calculation method: % Activity = (SB) * 100 / (PB). S = OD value of the inhibited group. P = OD value of the normal group, B = OD value of the PBS blank group (solvent background value). The results are shown in Table 1.

[0054] Table 1: Enzyme activity rates of the synthesized compounds AChE and BuChE at different concentrations according to the examples Given that most of the constructed compounds exhibited weak AChE inhibitory activity (less than 50% inhibition at 10 μM, significantly lower than positive controls such as donepezil, takrine, and galantamine), while a considerable number of compounds showed good BuChE inhibitory activity, we determined the enzyme activity of BuChE at multiple concentrations (1000, 200, 40, 8, 1.6, 0.32 nM) of these compounds and calculated the IC50 values ​​(Table 2). The IC50 calculation method (using the following formula to perform nonlinear fitting on the enzyme activity at different concentrations to calculate the IC50 value) is: %Activity = 100 * IC50 / (IC50 + [S]), where [S] represents the drug concentration.

[0055] Table 2: IC50 (nM) of some preferred compounds synthesized according to the examples for BuChE Example 12: In vitro toxicity test of the compound on mouse primary neurons This experiment aimed to evaluate the toxicity of some of the selected compounds (BuChE IC50 < 20 nm) to primary mouse neurons. Primary C57 mouse neurons were cultured in Neurobasal medium (containing 2% B27 and 1% GlutaMAX, neurons). Experimental groups included a normal neuronal control group (no compound), a blank control group (no cells, no compound), and a compound intervention group (50 μM target compound). Positive controls included the NMDA receptor antagonists cefotaxime and memantine, and common neuroprotective agents edaravone and butylphthalide. The specific procedure was as follows: ① Freshly extracted primary C57 mouse neurons were seeded in 96-well plates (neuron density 5 × 10⁻⁶). 4 ① Add 150 μL of the appropriate concentration of compound and 20 μL of the positive control (final concentration 50 μM) to each well, and incubate at 37℃ and 5% CO2 for 6-7 days, changing the medium every other day; ② Add the appropriate concentration of compound and 20 μL of positive control (final concentration 50 μM), and incubate at 37℃ and 5% CO2 for 24 hours; ④ Detect cell viability by MTT assay (add 10 μL of MTT working solution (5 mg / mL in DMSO) to each well under light-protected conditions, incubate at 5% CO2 for 3 hours, measure OD at 570 nm (reference 650 nm), and calculate according to the formula "viability (%) = [(experimental group OD - blank OD) / (normal control group OD - blank OD)] × 100%" (Table 3). Statistical analysis was performed using GraphPadPrism software, and experimental data are expressed as mean ± SD.

[0056] Table 3: Results of toxicity tests on primary mouse neurons of some of the preferred compounds (50 μM) synthesized according to the examples. Example 13: In vitro test of the protective effect of compounds against glutamate-damaged mouse primary neurons This experiment aimed to evaluate the protective effect of the above-selected compounds against glutamate-induced excitotoxicity in primary mouse neurons. Primary C57 mouse neurons were cultured in Neurobasal medium (containing 2% B27 and 1% GlutaMAX, neurons). Experimental groups included a normal neuronal control group (without glutamate / glycine and the compound), a blank control group (no cells, no compound), an injury group (treated with 100 μM glutamate + 10 μM glycine, culture medium prepared at pH 7.4), and a compound intervention group (100 μM glutamate + 10 μM glycine + target compound 5 / 0.5 μM). Positive controls included the NMDA receptor antagonists cyfortide and memantine, and common neuroprotective agents edaravone and butylphthalide. The specific procedure was as follows: ① Freshly extracted primary C57 mouse neurons were seeded in 96-well plates (neuron density 5 × 10⁶ cells / well). 4 ① Add 150 μL of the compound to each well and incubate at 37℃ and 5% CO2 for 6-7 days, changing the medium every other day; ② Add 20 μL of glutamic acid / glycine (final concentration 100 / 10 μM) and incubate at 37℃ and 5% CO2 for 30 min; ③ Add the corresponding concentration of the compound and 20 μL of the positive control (final concentration 5 / 0.5 μM) and incubate at 37℃ and 5% CO2 for 24 hours; ④ Detect cell viability using the MTT assay (add 10 μL of MTT working solution (5 mg / mL in DMSO) to each well under light-protected conditions, incubate at 5% CO2 for 3 hours, measure the OD at 570 nm (reference 650 nm), and calculate the survival rate (%) using the formula "(experimental group OD - blank OD) / (normal control group OD - blank OD)" × 100%. Statistical analysis was performed using GraphPad Prism software, and experimental data are expressed as mean ± SD.

[0057] Table 4: Protective effect (cell survival rate) of some preferred compounds synthesized according to the examples against glutamate-damaged nerve cells. In summary, the compound of general formula (I) of this invention exhibits highly selective inhibitory activity against butyrylcholinesterase (IC50). 50 These compounds exhibit low concentrations (down to sub-nanomolar levels) and show significant protective effects against glutamate-induced neuronal damage (increasing cell survival from 32% to 50-60% at a concentration of 5 μM). Furthermore, representative compounds at a concentration of 50 μM show no significant cytotoxicity to primary neurons (survival rate >80%). Therefore, these compounds can serve as lead molecules with both cholinesterase inhibition and neuroprotective functions for the development of drugs to treat neurological diseases such as Alzheimer's disease (AD).

Claims

1. A carbamate compound, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, enantiomer, diastereomer, racemate, or mixture thereof, characterized in that: The carbamate compound, or its pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, enantiomer, diastereomer, racemate, or mixture thereof, is shown in general formula (I): （I） In formula (Ⅰ), X is CH2, O, or S; n is 0 or 1; W1, W2, W3, W4 and W5 are each independently CH or N, and the total number of N does not exceed 2; R1 is a mono- or di-(C1-C4 alkyl) substituted amino group, or an unsubstituted or substituted 5- to 7-membered nitrogen heterocyclic group with 1-2 independent substituents selected from halogen, C1- to C4 alkyl, hydroxyl, hydroxymethyl, trifluoromethyl, carboxyl, C2- to C4 acyl, amide, etc., wherein the nitrogen heterocyclic group contains 0-1 additional heteroatom selected from O, S, N; R2 is selected from hydrogen, halogen, C1-C4 straight-chain or branched alkyl, C2-C4 straight-chain or branched alkenyl, C2-C4 straight-chain or branched alkynyl, C1-C4 alkoxy, C1-C4 alkylcarbonyloxy, cyano, nitro, hydroxy, hydroxymethyl, trifluoromethyl, carboxyl, mercapto, methyl mercapto, C1-C4 acyl, amide, aminosulfonyl, C1-C4 alkylsulfonyl, or two adjacent R2 substituents together with the carbon atom to which they are attached form a 5- to 7-membered ring; wherein the number of R2 is 0, 1, 2 or 3.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, enantiomer, diastereomer, racemate, or mixture thereof, characterized in that: The compounds are selected from the group consisting of: 。 3. Use of a carbamate compound as described in claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, enantiomer, diastereomer, racemate, or mixture thereof, characterized in that, The compound has selective inhibitory activity against butyrylcholinesterase, and the drug is used as a cholinesterase inhibitor for the treatment, prevention, or relief of Alzheimer's disease or functional impairments related to cholinergic system dysfunction.

4. Use of a carbamate compound as described in claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, enantiomer, diastereomer, racemate, or mixture thereof, characterized in that, The compound has selective inhibitory activity against butyrylcholinesterase and / or protective effects against glutamate-induced neuronal damage. The drug is used as a cholinesterase inhibitor and / or neuroprotective agent for the treatment, prevention, or relief of neurodegenerative diseases or nervous system disorders associated with cholinergic system dysfunction and / or neuronal damage.

5. A method for preparing a compound of general formula (I) as described in claim 1, characterized in that, Includes the following steps: S1. In a protic solvent and in the presence of an inorganic base, an amidine compound J is reacted with a β-keto ester compound Y to obtain a pyrimidine compound JY; the protic solvent is selected from one or more of methanol, ethanol, and isopropanol; the inorganic base is selected from one or more of potassium carbonate, sodium carbonate, cesium carbonate, sodium bicarbonate, sodium hydroxide, potassium hydroxide, potassium phosphate, sodium phosphate, dipotassium hydrogen phosphate, and disodium hydrogen phosphate. S2. In a polar aprotic solvent and in the presence of an organic base, a pyrimidine compound JY is reacted with a carbamoyl chloride compound d to obtain a carbamate compound JY-d; the polar aprotic solvent is selected from one or more of acetonitrile, tetrahydrofuran, dioxane, dichloromethane, chloroform, N,N-dimethylformamide, dimethyl sulfoxide, and N-methylpyrrolidone; the organic base is selected from one or more of triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, and pyridine. In the above reaction formulas, the definitions of R1, R2, X, W1-W4 substituents and n are as described in claim 1.

6. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises a therapeutically effective amount of one or more carbamate compounds as described in claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, enantiomer, diastereomer, racemate, or mixture thereof, and optionally a pharmaceutically acceptable carrier.

7. Use of the pharmaceutical composition as described in claim 6, characterized in that, The disease is selected from one or more of the following: Alzheimer's disease, Lewy body dementia, progressive supranuclear palsy, vascular dementia, myasthenia gravis, glaucoma, and cognitive or motor impairment caused by degenerative changes in the central or peripheral nervous system.

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