Arylalkylazole derivatives, processes for their preparation and use thereof

Abstract

The application belongs to the technical field of medicine, and particularly relates to an arylalkyl azole derivative, a preparation method and application thereof. The arylalkyl azole derivative is obtained by directly connecting 4H-1,2,4-triazole-4-yl to benz[d][1,3]thiazine as aryl through a chemical bond and introducing an alkoxy group at the C6 position of the aryl. The compound of the application has a novel structure, and as a GABA_A receptor agonist, exhibits excellent anticonvulsant activity in two kinds of epilepsy models of maximum electric shock and subcutaneous pentylenetetrazole. In particular, the compound exhibits an extremely high protection index PI>10 in the subcutaneous pentylenetetrazole model, indicating that the compound has the remarkable advantages of diverse action mechanism, wide therapeutic window and low neurotoxicity, and can be used for preparing an antiepileptic drug, and is particularly suitable for the treatment of refractory epilepsy.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to an arylalkylazole derivative, its preparation method, and its application. Background Technology

[0002] Epilepsy is a common chronic neurological disorder characterized by recurrent seizures caused by abnormal neuronal discharges. While existing antiepileptic drugs have some efficacy, 20%–30% of patients suffer from drug-resistant epilepsy, and long-term medication often results in adverse reactions such as central nervous system toxicity, drowsiness, and cognitive impairment. Clinical needs remain far from being met.

[0003] Gamma-aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the central nervous system, and enhancing GABAergic signal transduction is one of the important mechanisms of action of antiepileptic drugs. Arylalkylazole compounds, especially derivatives containing heterocyclic structures such as benzothiazole and triazole, have been reported to have GABA_A receptor regulatory activity, showing anticonvulsant potential.

[0004] However, existing arylalkylazole derivatives still suffer from insufficient activity, poor selectivity, low therapeutic index, or unsatisfactory blood-brain barrier permeability. For example, current research and development strategies mainly focus on improving activity in maximal electroshock models, while generally neglecting in-depth optimization of the subcutaneous pentylenetetrazol model, an important GABAergic mechanism model. This results in insufficient efficacy and safety of existing arylalkylazole derivatives in the subcutaneous pentylenetetrazol model. Summary of the Invention

[0005] To address the technical problem of insufficient effectiveness of existing arylalkylazole derivatives in subcutaneous pentylenetetrazol models, this invention provides a novel arylalkylazole derivative with strong anticonvulsant activity, high therapeutic index, and low neurotoxicity, along with its preparation method and applications.

[0006] This invention utilizes benzo[d][1,3]thiazide as a rigid hydrophobic core, directly linked to a 4H-1,2,4-triazol-4-yl group via chemical bonds, and introduces alkoxy chains of varying lengths at the C6 position of the aryl group to obtain arylalkylazole derivatives. These arylalkylazole derivatives enhance the regulatory activity on the GABAergic system, particularly strengthening the anticonvulsant effect in the scPTZ model; furthermore, they improve the therapeutic index, reduce neurotoxicity, and optimize oral bioavailability and blood-brain barrier permeability, meeting clinical drug requirements.

[0007] This invention directly couples the rigid plane of benzothiazide with the bioactive characteristics of the triazole ring, and systematically regulates the molecule's lipid-water partition coefficient, conformational flexibility, and metabolic stability by precisely introducing alkoxy side chains of different lengths. The aim is to optimize its ability to cross the blood-brain barrier and its in vivo pharmacokinetic properties. This structural design is intended to enable it to act efficiently on GABA_A receptors, not only directly enhancing receptor function as a positive allosteric modulator, but also potentially achieving optimal interaction with the receptor's hydrophobic cavity through the flexible regulation of the side chains, thereby achieving a potent and safe antiepileptic effect.

[0008] The first objective of this invention is to provide an arylalkylazole derivative, the chemical structural formula of which is as follows:

[0009] ;

[0010] In the formula, R represents C1 to C2. 10 Alkyl groups.

[0011] Preferably, the core skeleton of the arylalkylazole derivative is linked as follows: the benzo[d][1,3]thiazine ring and the 1,2,4-triazole ring are directly connected by a CN single bond without any spacer atoms; the molecular conformation is as follows: the substituent R side chain exhibits an extended conformation in the crystal and participates in intermolecular stacking.

[0012] Preferably, R is -C7H 15 .

[0013] Preferably, the arylalkylazole derivative is 6-(heptoxy)-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazine, with the following structural formula:

[0014] .

[0015] A second objective of this invention is to provide a method for preparing arylalkylazole derivatives, comprising the following steps:

[0016] Intermediate 1 undergoes a cyclization reaction with (E)-N'-[(E)-(dimethylamino)methylene]-N,N-dimethylformylhydrazine formamide under a catalyst to give intermediate 2; intermediate 2 undergoes a demethylation reaction with boron tribromide to give intermediate 3; under basic conditions, intermediate 3 undergoes a nucleophilic substitution reaction with a haloalkane in a solvent to give an arylalkylazole derivative; the specific synthetic route is as follows:

[0017] .

[0018] Preferably, the molar ratio of intermediate 1 to (E)-N'-[(E)-(dimethylamino)methylene]-N,N-dimethylformylhydrazine formamide is 1:1 to 2; the molar ratio of intermediate 2 to boron tribromide is 1:2; and the molar ratio of intermediate 3 to haloalkane is 1:3 to 5. The higher the proportion of haloalkane, the higher the yield and the faster the reaction rate.

[0019] Preferably, the haloalkane is n-heptane bromo, ethane bromo, propane bromo, butane bromo, isopentane bromo, 2-methylbutane bromo, 4-methylpentane bromo, 5-methylhexane bromo, n-hexane bromo, n-octane bromo, or n-decane bromo.

[0020] Preferably, the solvent is at least one of acetonitrile, acetone and N,N-dimethylformamide; and the base is potassium carbonate.

[0021] Preferably, the specific preparation method of intermediate 2 is as follows:

[0022] Intermediate 1 was mixed with (E)-N'-[(E)-(dimethylamino)methylene]-N,N-dimethylformylhydrazine formamide, and a catalyst was added under ice bath conditions. The mixture underwent a cyclization reaction at 135℃ to 145℃ to obtain intermediate 2. The catalyst was p-toluenesulfonic acid. The cyclization reaction took 4 to 6 hours.

[0023] Preferably, the specific preparation method of intermediate 3 is as follows:

[0024] Intermediate 2 was mixed with a solvent, and boron tribromide solution was added at 0°C. A demethylation reaction was then carried out at room temperature to obtain intermediate 3. The solvent was dichloromethane, and the demethylation reaction time was 4 to 6 hours.

[0025] A third objective of this invention is to provide the application of arylalkylazole derivatives in the preparation of drugs for the prevention and treatment of chronic nervous system diseases.

[0026] Preferably, the active ingredient of the drug for preventing and treating chronic nervous system diseases is an arylalkylazole derivative, its isomer, or a pharmaceutically acceptable salt thereof.

[0027] Preferably, pharmaceutically acceptable salts are salts formed from arylalkylazole derivatives with acids or arylalkylazole derivatives with bases.

[0028] Preferred, pharmaceutically acceptable salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, phosphate, maleate, fumarate, citrate, tartrate, sodium, potassium, and calcium salts.

[0029] Preferred chronic neurological disease is epilepsy.

[0030] Preferably, the epilepsy is refractory epilepsy.

[0031] This drug composition inhibits various seizure models, including maximal electroshock (MES), subcutaneous pentylenetetrazol (PTZ), and 3-mercaptopropionic acid-induced seizures (3-MP), by regulating GABA_A receptor function. It also reduces seizure intensity and frequency in the chronic PTZ ignition model (simulating refractory epilepsy), demonstrating broad-spectrum antiepileptic activity.

[0032] Compared with the prior art, the present invention has the following technical effects:

[0033] 1. This invention directly couples the rigid plane of benzothiazide with the bioactive characteristics of the triazole ring, and systematically regulates the molecule's lipid-water partition coefficient, conformational flexibility, and metabolic stability by introducing alkoxy side chains of different lengths at the C6 position of benzothiazide. The aim is to optimize its ability to cross the blood-brain barrier and its in vivo pharmacokinetic properties. This structural design is intended to enable it to act efficiently on GABA_A receptors, not only directly enhancing receptor function as a positive allosteric modulator, but also potentially achieving optimal interaction with the receptor's hydrophobic cavity through the flexible regulation of the side chains, thereby achieving a potent and safe antiepileptic effect.

[0034] 2. Compared with benzothiazide derivatives in the prior art that overly rely on the MES model and generally exhibit poor activity in the PTZ model, the arylalkylazole derivatives of this invention show potent activity in both MES and PTZ epilepsy models with different mechanisms; especially in the PTZ model of ED 50 With a concentration as low as 8.86 mg / kg, it is significantly superior to most similar derivatives in the prior art. This indicates that the arylalkylazole derivatives of this invention have a strong positive regulatory effect on the GABAergic system, and can effectively combat seizures caused by insufficient GABAergic inhibition. It is expected to have therapeutic effects on epilepsy types that are insensitive to existing MES-guided drugs, and solves the problem of "insufficient effectiveness in PTZ models" in the prior art.

[0035] 3. The arylalkylazole derivatives of this invention exhibit potent anti-subcutaneous pentylenetetrazol activity while significantly reducing neurotoxicity. The median neurotoxicity dose (LD50) of compound H-10 is 93.67 mg / kg, resulting in a protection index as high as 10.6. This protection index is significantly higher than that of most AAA derivatives reported in the prior art and first-line clinical drugs such as carbamazepine. This demonstrates that the arylalkylazole derivatives of this invention have an extremely broad therapeutic window and excellent drug safety for GABAergic system-related epilepsy types, overcoming the shortcomings of "low protection index" and "high activity accompanied by high toxicity" in the prior art, and significantly improving the expected clinical safety.

[0036] 4. Mechanistic studies of the present invention confirm that arylalkylazole derivatives not only directly enhance GABA_A receptor function as positive allosteric regulators, but also induce long-term adaptive changes such as downregulation of ABAT and GAT1 expression after long-term administration. This means that these arylalkylazole derivatives can synergistically enhance GABAergic inhibitory efficacy from two levels: "enhancing inhibitory signal generation" and "reducing inhibitory signal clearance." This multi-target, multi-level synergistic mechanism, compared with compounds with relatively broad or single mechanisms in the prior art, can bring superior efficacy and lower risk of drug resistance, and is especially suitable for the treatment of refractory epilepsy. Attached Figure Description

[0037] Figure 1 The molecular structure diagram of the H10 compound of Example 1, determined by X-ray single-crystal diffraction.

[0038] Figure 2 The diagram shows the molecular docking results of the H10 compound and the 6HUP protein in Example 1; where (a) is the molecular docking result of the H10 compound and the 6HUP protein in Example 1, and (b) is an enlarged view of (a).

[0039] Figure 3 The original diagram shows the effect of the H10 compound synthesized in Example 1 on the expression of GABAergic system-related proteins.

[0040] Figure 4 The curves show the changes in the protective rate of compound H10 against scPTZ-induced seizures at different time points after oral administration.

[0041] Figure 5 The effect of compound H10 synthesized in Example 1 on seizure score and duration in a chronic ignition model is shown. (a) represents the seizure score in the pre-ignition experiment, (b) represents the seizure latency in the pre-ignition experiment, and (c) represents the duration of epileptic clonic episodes in the pre-ignition experiment.

[0042] Figure 6 The effect of compound H10 synthesized in Example 1 on seizure score and duration in a chronic ignition model is shown. (a) represents the percentage of seizure grade scores in the post-ignition experiment, (b) represents the duration of epileptic clonic seizures in the post-ignition experiment, (c) represents the seizure grade scores in the post-treatment experiment, and (d) represents the duration of epileptic clonic seizures in the post-treatment experiment.

[0043] Figure 7 HE staining of compound H-10 synthesized in Example 1 for pre-ignition experiments in the PTZ ignition model.

[0044] Figure 8HE staining of compound H-10 synthesized in Example 1 in a post-treatment experiment in a PTZ ignition model.

[0045] Figure 9 HE staining of the recovered PTZ ignition model using compound H-10 synthesized in Example 1. Detailed Implementation

[0046] It should be noted that benzothiazide is a nitrogen- and sulfur-containing benzo[a]heterocyclic compound. Its unique rigid planar structure and the hydrophobic interactions and potential hydrogen bonding capabilities provided by the sulfur atoms make it a superior framework in medicinal chemistry, especially in the design of drugs for the central nervous system. This structure can stably bind to a variety of biological targets through π-π stacking and hydrophobic interactions. This characteristic makes it frequently used to construct molecules with neuroactive properties. In the field of antiepileptic drug research, the benzothiazide framework has shown clear application value. Some molecular simulation studies further support the targeting potential of this framework, confirming that derivatives containing the benzothiazide structure can effectively bind to the benzodiazepine site of the GABA_A receptor, providing a structural biology explanation for its anticonvulsant effect. Triazole heterocycles, especially 1,2,4-triazoles, are classic "privileged fragments" used in medicinal chemistry to optimize molecular properties. This structure has several advantages: the nitrogen atom in the triazole ring can act as a hydrogen bond acceptor, enhancing the affinity for biological targets; moreover, its moderate lipophilicity helps balance the lipid-water partition coefficient of the molecule, which is crucial for improving the drug's ability to cross the blood-brain barrier. It has affinity for some targets related to epilepsy treatment and exhibits highly selective and low-toxicity effective anticonvulsant activity.

[0047] Based on this, the present invention uses benzo[d][1,3]thiazine Ar as a rigid hydrophobic core, directly linked to the "privileged fragment" 4H-1,2,4-triazol-4-yl Z through chemical bond L, and introduces alkoxy chains of different lengths at the C6 position of the aryl group. While optimizing the lipid-water partition coefficient, it effectively regulates the molecular conformational flexibility, hydrophobic interaction area and metabolic stability, achieving multi-parameter synergistic optimization and providing an advantageous structural framework for antiepileptic drugs.

[0048] To enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings.

[0049] Unless otherwise specified, all reagents used in this invention are commercially available, and all methods used are conventional techniques in the art.

[0050] Dimethyl sulfoxide (DMSO) is the English name for dimethyl sulfoxide; pentylenetetrazole (PTZ) is the English name for pentylenetetrazole.

[0051] The synthetic route of intermediate 1 used in the following embodiments is as follows:

[0052] .

[0053] The specific synthesis method is as follows:

[0054] Step 1, Synthesis of (2-amino-5-methoxyphenyl)methanol:

[0055] In a 1L three-necked round-bottom flask equipped with a nitrogen balloon, a constant-pressure dropping funnel, and a thermometer, tetrahydrofuran (250 mL) and methyl 2-amino-5-methoxybenzoate (45 g, 248.36 mmol, 1.0 eq) were added. The reaction system was cooled to 0°C, and 149.02 mL of 2.5 M lithium aluminum hydride solution was slowly added dropwise, with tetrahydrofuran as the solvent. After the addition was complete, the temperature was raised to 20°C and the reaction mixture was stirred for 16 h. The reaction progress was monitored by thin-layer chromatography, using petroleum ether and ethyl acetate as the developing solvents in a 1:1 volume ratio. After the starting material spot disappeared, the reaction solution was cooled to 0°C–5°C, and water (15 mL) was slowly added, followed by dropwise addition of 15% sodium hydroxide aqueous solution (15 mL), and then slow addition of water (45 mL). The reaction mixture was extracted with dichloromethane, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain intermediate 0, namely (2-amino-5-methoxyphenyl)methanol.

[0056] The properties of (2-amino-5-methoxyphenyl)methanol are as follows:

[0057] Black solid, 65 g, yield 85.43%, melting point 64.28℃~65.55℃. 1H NMR data are as follows: 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 6.73 (d, J = 2.2 Hz, 1H), 6.58 (d, J = 3.5 Hz, 2H), 5.11–4.44 (m, 3H), 4.35 (s, 2H), 3.63 (s, 3H); ¹H NMR data are as follows: 13 C NMR (101 MHz, DMSO-d6) δ 151.31, 139.94, 127.53, 116.06, 113.69, 113.34, 61.25, 55.72; NMR mass spectrometry data are ESI-MS: m / z 136.0758 [M+H] + .

[0058] Step 2, Synthesis of 6-methoxy-4H-benzo[d][1,3]thiazine-2-ammonium:

[0059] Intermediate 0 (65 g, 424.34 mmol, 1.0 eq) was placed in a 1 L three-necked flask, and concentrated hydrochloric acid (300 mL) was added. The mixture was stirred at 100 °C for 15 min. The reaction solution was concentrated to obtain a residue. The nitrogen balloon and thermometer were reattached, and propylene glycol (600 mL) was added. Thiourea (35.53 g, 466.78 mmol, 1.1 eq) was added at 0 °C, and the mixture was stirred at 85 °C for 16 h. The reaction progress was monitored by thin-layer chromatography, with petroleum ether and ethyl acetate as the developing solvents in a 1:1 volume ratio. Water (2 L) was added to the reaction system, and the pH of the aqueous phase was adjusted to 4 with 2 M HCl (approximately 100 mL). The mixture was stirred at room temperature for 15 min and extracted with dichloromethane. All aqueous phases were combined, and the pH was adjusted to 10 with 2 M NaOH (200 mL). The mixture was stirred at 0 °C for 15 min, and a solid precipitated. The solid was filtered, and the filter cake was washed three times with water and petroleum ether, respectively. The obtained solid was purified by silica gel column chromatography (300-400 mesh) using dichloromethane / methanol (50:1) as eluent to obtain intermediate 1, namely 6-methoxy-4H-benzo[d][1,3]thiazine-2-ammonium.

[0060] The properties of 6-methoxy-4H-benzo[d][1,3]thiazine-2-ammonium are as follows:

[0061] Yellow solid, 30.0 g, yield 29.48%, melting point 186.99℃~187.7℃. 1H NMR data are as follows: 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 6.80 (d, J = 9.4 Hz, 2H), 6.75 (dd, J = 5.7, 2.8 Hz, 3H), 3.87 (s, 2H), 3.70 (s, 3H); ¹H NMR data are as follows: 13 C NMR (101 MHz, DMSO-d6) δ 155.28, 152.89, 140.29, 124.39, 120.92, 113.76, 112.48, 55.68, 29.13; NMR mass spectrometry data are ESI-MS: m / z 195.0596 [M+H] + .

[0062] Example 1

[0063] A method for preparing an arylalkylazole derivative includes the following steps:

[0064] Step 1, Synthesis of 6-methoxy-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide:

[0065] In a 500 mL three-necked flask equipped with a nitrogen balloon and a thermometer, 120 mL of o-xylene, intermediate 1 (30 g, 154.44 mmol, 1.0 eq), and (E)-N'-[(E)-(dimethylamino)methylene]-N,N-dimethylformylhydrazine formamide (26.35 g, 185.33 mmol, 1.2 eq) were added. p-Toluenesulfonic acid (2.5 g, 14.52 mmol, 0.094 eq) was added at 0 °C, followed by stirring at 140 °C for 4 h. The reaction progress was monitored by thin-layer chromatography using petroleum ether and ethyl acetate as the developing solvent (volume ratio 1:1). The mixture was cooled to 20 °C, and the reaction was quenched by adding 200 mL of petroleum ether. The solid was collected by filtration. The crude product was purified by silica gel column chromatography (300-400 mesh) using dichloromethane / methanol (60:1) as eluent to obtain intermediate 3, namely 6-methoxy-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide.

[0066] The properties of 6-methoxy-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide are as follows:

[0067] Yellow solid, 8.15 g, yield 19.64%, melting point 150.34℃~150.97℃. 1H NMR data are as follows: 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 9.16 (s, 2H), 7.23 (d, J = 8.5 Hz, 1H), 6.99 – 6.93 (m, 2H), 4.33 (s, 2H), 3.80 (s, 3H); ¹H NMR data are as follows: 13 C NMR (101 MHz, DMSO-d6) δ 159.52, 143.21, 141.16, 135.69, 127.82, 122.02, 114.38, 113.16, 55.99, 29.15; NMR mass spectrometry data are ESI-MS: m / z 269.0474 [M+Na] + .

[0068] Step 2, Synthesis of 2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazine-6-ol:

[0069] Dichloromethane (100 mL) and intermediate 2 (9.5 g, 38.57 mmol, 1.0 eq) were added to a 250 mL three-necked flask equipped with a nitrogen balloon, dropping funnel, thermometer, and tail gas absorption device. 38.57 mL of 2 M boron tribromide solution (DCM) was slowly added dropwise at 0 °C over approximately 1 hour. The mixture was then brought to room temperature and stirred for 6 hours. The reaction was monitored by TLC until completion. The reaction mixture was poured into an ice-cold saturated sodium bicarbonate solution (300 mL), filtered, and the resulting filter cake was washed with water (100 mL) for 30 minutes with stirring. This process was repeated twice. The solid was then washed again with dichloromethane (100 mL) at room temperature for 30 minutes, and the mixture was filtered. The obtained solid was washed successively with dichloromethane and petroleum ether, and finally stirred with dichloromethane / methanol (3:1) for 30 min, filtered, and the filter cake was washed with petroleum ether to obtain intermediate 3, namely 2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazin-6-ol.

[0070] The properties of 2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazine-6-ol are as follows:

[0071] Yellow solid, melting point 186.32℃~186.91℃. 1H NMR data are as follows: 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 9.85 (s, 1H), 9.14 (s, 2H), 7.13 (d, J = 8.5 Hz, 1H), 6.79 (dd, J = 8.5, 2.5 Hz, 1H), 6.72 (d, J = 2.4 Hz, 1H), 4.28 (s, 2H); ¹H NMR data are as follows: 13 C NMR (101 MHz, DMSO-d6) δ 158.22, 141.92, 141.39, 134.17, 128.06, 121.93, 115.74, 114.39, 29.12; NMR mass spectrometry data are ESI-MS: m / z 233.04922 [M+H] + .

[0072] Step 5: Synthesis of arylalkylazole derivatives:

[0073] 0.2155 mmol of intermediate 3, 0.6512 mmol of potassium carbonate, and 25 mL of acetonitrile were added to a 100 mL round-bottom flask. 0.6465 mmol of n-heptane bromide was added under stirring. The reaction mixture was refluxed at 80 °C for 6 hours. After the reaction was complete, the mixture was cooled to room temperature, and the solvent was removed by vacuum distillation. The residue was extracted with dichloromethane, and the organic phases were combined and dried over anhydrous sodium sulfate. The mixture was filtered and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using dichloromethane and methanol as eluents in a volume ratio of 20:1, yielding a white solid 6-(heptoxy)-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide, an arylalkylazole derivative, in 65% yield, denoted as H-10.

[0074] The properties of 6-heptoxy-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide are as follows:

[0075] White solid, 43.94 mg, yield 61.7%, melting point 97.88℃~98.03℃. ¹H NMR (400 MHz, DMSO-d⁶) δ 9.16 (s, 2H), 7.21 (d, J = 8.4 Hz, 1H), 6.96–6.92 (m, 2H), 4.32 (s, 2H), 3.99 (t, J = 6.5 Hz, 2H), 1.73–1.68 (m, 2H), 1.42–1.37 (m, 2H), 1.27 (s, 6H), 0.86 (t, J = 6.8 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d⁶) δ 158.95, 143.10, 141.17, 135.53, 127.79, 121.99, 114.81, 113.61, 68.30, 31.71, 29.11, 29.09, 28.89, 25.92, 22.53, 14.42; ESI-MS: m / z 331.15861 [M+H]+, melting point 97.8℃-98.0℃.

[0076] Example 2

[0077] A method for preparing an arylalkylazole derivative is basically the same as that in Example 1, except that the alkyl halide is different.

[0078] The haloalkanes are bromoethane, bromopropane, bromobutane, bromoisopentane, bromo2-methylbutane, bromo4-methylpentane, bromo5-methylhexane, bromon-hexane, bromon-octane, or bromon-decane.

[0079] The synthesized arylalkylazole derivatives are as follows:

[0080] The haloalkane is bromoethane, yielding 6-ethoxy-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazine, with the following properties:

[0081] Yellow solid, 15.40 mg, yield 27.45%, melting point 171.30℃~172.63℃. 1H NMR data are as follows: 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 9.16 (s, 2H), 7.22 (d, J = 8.4 Hz, 1H), 6.97–6.92 (m, 2H), 4.32 (s, 2H), 4.06 (q, J = 7.0 Hz, 2H), 1.34 (t, J = 6.9 Hz, 3H); ¹³C NMR data are as follows: 13 C NMR (101 MHz, DMSO-d6) δ 158.78, 143.13, 141.17, 135.55, 127.81, 122.00, 114.82, 113.57, 63.94, 29.13, 15.07; NMR mass spectrometry data are ESI-MS: m / z 261.08037 [M+H] + .

[0082] The haloalkane was bromopropane, and 6-propoxy-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide was synthesized, with the following properties:

[0083] White solid, 24.30 mg, yield 41.10%, melting point 155.94℃~157.93℃. 1H NMR data are as follows: 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 9.16 (s, 2H), 7.22 (d, J = 8.5 Hz, 1H), 6.96 (dd, J = 12.0, 3.4 Hz, 2H), 4.33 (s, 2H), 3.97 (t, J = 6.5 Hz, 2H), 1.74 (h, J = 7.1 Hz, 2H), 0.98 (t, J = 7.4 Hz, 3H); ¹³C NMR data are as follows: 13C NMR (101 MHz, DMSO-d6) δ 158.98, 143.13, 141.18, 135.58, 127.83, 122.02, 114.88, 113.68, 69.84, 39.72, 29.13, 22.46, 10.81; NMR mass spectrometry data are ESI-MS: m / z 275.09593 [M+H] + .

[0084] The haloalkane was bromobutane, and 6-butoxy-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide was synthesized, with the following properties:

[0085] White solid, 25.8 mg, yield 41.52%, melting point 140.62℃~141.52℃. 1H NMR data are as follows: 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 9.16 (s, 2H), 7.21 (d, J = 8.4 Hz, 1H), 6.98 - 6.92 (m, 2H), 4.32 (s, 2H), 4.00 (t, J = 6.4 Hz, 2H), 1.70 (p, J = 6.6 Hz, 2H), 1.47 - 1.40 (m, 2H), 0.93 (t, J = 7.4 Hz, 3H); ¹³C NMR data are as follows: 13 C NMR (101 MHz, DMSO-d6) δ 158.96, 143.11, 141.17, 135.55, 127.80, 122.00, 114.84, 113.63, 68.03, 31.14, 29.12, 19.17, 14.13; NMR mass spectrometry data are ESI-MS: m / z 289.11146 [M+H]+.

[0086] The haloalkane was bromoisopentane, and 6-isopentoxy-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazine was synthesized, with the following properties:

[0087] White solid, 41.43 mg, yield 27.00%, melting point 136.9℃~137.24℃. The 1H NMR data are as follows: 1H NMR (400 MHz, DMSO-d6) δ 9.16 (s, 2H), 7.21 (d, J = 8.4 Hz, 1H), 6.99 - 6.94 (m, 2H), 4.33 (s, 2H), 4.03 (t, J = 6.6 Hz, 2H), 1.78 (dt, J = 13.3, 6.7 Hz, 1H), 1.62 (q, J = 6.7 Hz, 2H), 0.93 (d, J = 6.6 Hz, 6H); the 13C NMR data are as follows: 101 MHz, DMSO-d6 δ 158.95, 143.11, 141.17, 135.56, 127.80, 121.99, 114.85, 113.64, 66.81, 37.83, 29.12, 25.08, 22.87; NMR mass spectrometry data are ESI-MS: m / z 303.12722 [M+H] + .

[0088] The haloalkane is bromo-2-methylbutane. The properties of the synthesized 6-(2-methylbutoxy)-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide are as follows:

[0089] White solid, 47.72 mg, yield 31.10%, melting point 134.83℃~135.59℃. 1H NMR data are as follows: 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 9.17 (s, 2H), 7.21 (d, J = 8.2 Hz, 1H), 6.96 (d, J = 8.8 Hz, 2H), 4.32 (s, 2H), 3.89 - 3.84 (m, 1H), 3.81 - 3.76 (m, 1H), 1.81 (dq, J = 12.9, 6.4 Hz, 1H), 1.23 (dt, J = 13.8, 7.7 Hz, 2H), 0.96 (d, J = 6.7 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H); ¹³C NMR data are as follows: 13C NMR (101 MHz, DMSO-d6) δ 159.10, 143.11, 141.17, 135.55, 127.80, 122.00, 114.86, 113.70, 73.07, 34.49, 29.10, 25.99, 16.72, 11.56; ESI-MS; NMR mass spectrometry data is m / z 303.12716 [M+H] + .

[0090] The haloalkane was bromo-4-methylpentane, and 6-((4-methylpentyl)oxy)-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide was synthesized, with the following properties:

[0091] White solid, 28.89 mg, yield 9.70%, melting point 130.29℃~131.13℃. 1H NMR data are as follows: 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 9.17 (s, 2H), 7.22 (d, J = 8.5 Hz, 1H), 6.98 - 6.93 (m, 2H), 4.33 (s, 2H), 3.99 (t, J = 6.5 Hz, 2H), 1.75 - 1.69 (m, 2H), 1.58 (dt, J = 13.3, 6.7 Hz, 1H), 1.33 - 1.28 (m, 2H), 0.90 (s, 3H), 0.88 (s, 3H); ¹³C NMR data are as follows: 13 C NMR (101 MHz, DMSO-d6) δ 158.95, 143.13, 141.19, 135.55, 127.80, 122.02, 114.81, 113.64, 68.60, 35.12, 29.10, 27.72, 27.02, 22.93; NMR mass spectrometry data are ESI-MS: m / z 317.14289 [M+H] + .

[0092] The haloalkane was bromo-5-methylhexane. 6-((5-methylhexyl)oxy)-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazine was synthesized, and its properties are as follows:

[0093] White solid, 20.20 mg, yield 28.37%, melting point 83.24℃~84.01℃. 1H NMR data are as follows: 1¹H NMR (400 MHz, DMSO-d⁶) δ 9.17 (s, 2H), 7.21 (d, J = 8.4 Hz, 1H), 6.95 (dd, J = 10.8, 2.1 Hz, 2H), 4.32 (s, 2H), 3.99 (t, J = 6.5 Hz, 2H), 1.70 (p, J = 6.7 Hz, 2H), 1.53 (dt, J = 13.3, 6.6 Hz, 1H), 1.40 (dt, J = 15.5, 7.5 Hz, 2H), 1.24 - 1.19 (m, 2H), 0.87 (s, 3H), 0.85 (s, 3H); ¹H NMR data are as follows: 13 C NMR (101MHz, DMSO-d6) δ 158.95, 143.10, 141.17, 135.54, 127.80, 122.00, 114.81, 113.62, 68.31, 38.56, 29.33, 29.11, 27.88, 23.75, 22.94; NMR mass spectrometry data are ESI-MS: m / z 331.15845 [M+H] + .

[0094] The haloalkane was n-hexane bromo, and 6-hexyloxy-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide was synthesized, with the following properties:

[0095] White solid, 17.30 mg, yield 25.37%, melting point 87.34℃~88.29℃. 1H NMR data are as follows: 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 9.16 (s, 2H), 7.22 (d, J = 8.4 Hz, 1H), 6.98 - 6.93 (m, 2H), 4.33 (s, 2H), 4.00 (t, J = 6.5 Hz, 2H), 1.71 (q, J = 6.9 Hz, 2H), 1.45 -1.39 (m, 2H), 1.31 (h, J = 3.8 Hz, 4H), 0.90 – 0.86 (m, 3H); ¹H NMR data are as follows: 13CNMR (101 MHz, DMSO-d6) δ 158.96, 143.12, 141.17, 135.55, 127.80, 122.01, 114.84, 113.64, 68.33, 31.43, 29.11, 29.04, 25.61, 22.54, 14.37; NMR-MS data are ESI-MS: m / z 317.14300 [M+H] + .

[0096] The haloalkane was bromooctane, and 6-octoxy-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide was synthesized, with the following properties:

[0097] White solid, 21.60 mg, yield 29.97%, melting point 91.41℃~92.69℃. 1H NMR data are as follows: 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 9.16 (s, 2H), 7.21 (d, J = 8.4 Hz, 1H), 6.98 - 6.91 (m, 2H), 4.32 (s, 2H), 3.99 (t, J = 6.5 Hz, 2H), 1.71 (p, J = 6.6 Hz, 2H), 1.45 -1.36 (m, 2H), 1.27 (d, J = 10.3 Hz, 8H), 0.85 (d, J = 7.0 Hz, 2H); ¹H NMR data are as follows: 13 C NMR (101 MHz, DMSO-d6) δ 158.98, 143.08, 141.14, 135.56, 127.81, 121.97, 114.85, 113.66, 68.35, 31.70, 29.17, 29.13, 29.12, 29.08, 25.95, 22.54, 14.40; NMR mass spectrometry data are ESI-MS: m / z 345.1753 [M+H] + .

[0098] The haloalkane was bromodecane, and 6-decoxy-2-(4H-1,2,4-triazol-4-yl)-4H-benzo[d][1,3]thiazide was synthesized, with the following properties:

[0099] White solid, 26.40 mg, yield 32.88%, melting point 93.77℃~95.18℃. 1H NMR data are as follows: 1¹H NMR (400 MHz, DMSO-d⁶) δ 9.16 (s, 2H), 7.21 (d, J = 8.4 Hz, 1H), 6.97 - 6.92 (m, 2H), 4.32 (s, 2H), 3.98 (t, J = 6.5 Hz, 2H), 1.70 (q, J = 6.9 Hz, 2H), 1.43 -1.37 (m, 2H), 1.24 (s, 12H), 0.86 - 0.83 (m, 3H); ¹H NMR data are as follows: 13 C NMR (101MHz, DMSO-d6) δ 158.95, 143.08, 141.15, 135.53, 127.79, 121.98, 114.80, 113.61, 68.30, 31.77, 29.49, 29.43, 29.22, 29.18, 29.11, 29.08, 25.94, 22.57, 14.41; NMR mass spectrometry data are ESI-MS: m / z 373.20561 [M+H] + .

[0100] Structure and purity testing.

[0101] 1. Melting point test.

[0102] Using the capillary method, referring to Method I of General Chapter 0612 of the 2020 edition of the Chinese Pharmacopoeia, the melting point of compound H-10 was determined to be 97.8℃~98℃ (uncorrected), and the melting range did not exceed 1℃; thus, it is shown that the H-10 prepared in Example 1 has high purity.

[0103] 2. Purity test.

[0104] High-performance liquid chromatography (HPLC) analysis was performed, with the following HPLC conditions: C18 column, mobile phase A being water and mobile phase B being acetonitrile, with a volume ratio of A to B of 90:10, and detection wavelength of 254 nm.

[0105] The chemical purity of compound H-10 was measured to be ≥98.0%, which meets the requirements for compound purity in pharmacological studies.

[0106] 3. Crystal structure testing.

[0107] Compound H-10 was cultured via slow solvent evaporation to obtain colorless bulk single crystals suitable for X-ray single-crystal diffraction analysis. Diffraction data were collected using Cu Kα radiation at a low temperature of 145 K on a Bruker D8 VENTURE diffractometer; λ = 1.54178 Å. The crystal structure was determined by the direct method (SHELXT) and based on the full-matrix least squares method using F... 2 Fine-tuning was performed; crystallographic data are shown in Table 1.

[0108] Table 1 Crystallographic data of H-10

[0109]

[0110] like Figure 1 As shown, the absolute three-dimensional molecular structure of H-10 was determined by X-ray single-crystal diffraction analysis. The final refinement deviation factors R1=0.0470 and wR2=0.1432 are within the generally accepted excellent range, fully demonstrating the accuracy and reliability of the crystal structure model. Crystal diffraction shows that the structure is a bihydrate of the target structure, and the formed twin form proves the correctness of the target compound structure.

[0111] The crystal structure clearly shows that the core framework is connected by a benzo[d][1,3]thiazine ring and a 1,2,4-triazole ring directly linked by a CN single bond (the connecting chain L is a chemical bond), with a bond length of 1.345(2) Å. This value is completely consistent with the theoretical expectation, confirming the core structure of the arylalkylazole derivative prepared in this embodiment of the invention at the atomic level. The molecular conformation is as follows: the heptoxy side chain exhibits an extended conformation in the crystal and participates in intermolecular stacking. The molecular state in the crystalline state is as follows: each unit contains two target molecules, forming a clear dimer structure through weak interactions such as intermolecular hydrogen bonds. This provides a key structural basis for understanding the solid-state properties and crystal form characteristics of the compound.

[0112] Pharmacodynamic testing.

[0113] 1. Evaluation of anticonvulsant activity.

[0114] Twenty-four hours prior to the experiment, mice were screened by stimulating their ears with 110V, 50Hz AC current for 0.2 seconds. Mice exhibiting hind limb rigidity were eligible for the experiment. During the experiment, 30 minutes after intraperitoneal injection of compound H-10, or after oral administration reaching peak concentration, mice were stimulated with 110V, 50Hz AC current. Mice that did not exhibit hind limb rigidity were considered positive, indicating that compound H-10 has anticonvulsant activity, providing a preliminary evaluation of the efficacy of the unknown drug. This anticonvulsant experimental model is designated as the MES model.

[0115] Table 2 Anticonvulsant test model

[0116]

[0117] 2. Pentyltetrazole-induced seizure test.

[0118] Grouping: Mice were randomly divided into three groups: blank group, positive control group and experimental group, with 10 mice in each group; the blank group was denoted as DMSO.

[0119] Drug administration and modeling:

[0120] Acute ignition model: As shown in Table 3, mice in the experimental group were given compound H-10, and the positive control group was given carbamazepine, all administered intraperitoneally. The blank control group received no treatment. After 30 minutes, except for the blank control group, the positive control group and the experimental group were given PTZ subcutaneously at a dose of 100 mg / kg. Then, according to the Racine criteria for grade 6 seizures in neurology (as shown in Table 4), the mice were observed behaviorally. Observation continued for 0.5 hours, and relevant parameters and indicators during seizures were observed and recorded. Recorded items included the latency and duration of clonic seizures, seizure behavior grade, and the percentage of mortality after the seizure (as shown in Table 3). This pentylenetetrazole-induced seizure model is designated as the PTZ model.

[0121] Table 3. Pentylenetetrazole-induced seizure test models

[0122]

[0123] Note: " / " indicates that no drug treatment was added.

[0124] Table 4 Neurological criteria for grade 6 epileptic seizures

[0125]

[0126] 3. Seizure-inducing test using the 3-mercaptopropionic acid model:

[0127] The method was the same as the pentylenetetrazol-induced seizure test, except that 3-mercaptopropionic acid was administered intraperitoneally at a dose of 75 mg / kg 30 minutes later; the purpose was to determine the agonistic effect of H-10 on GAB synthase GAD. The latency and duration of clonic seizures, seizure behavior grade, and post-ictal mortality rate were recorded, and the results are shown in Table 5.

[0128] Statistical analysis: SPSS 19 statistical software was used to analyze the above observations. The data were then processed and analyzed. Quantitative data in this experimental analysis are expressed as mean ± standard deviation (x ± s). Comparison of means among multiple samples and pairwise comparisons of means were performed using analysis of variance. Ordinal data were analyzed using nonparametric tests. This 3-mercaptopropionic acid model for inducing seizures is denoted as the 3-MP model.

[0129] Table 5. 3-Mercaptopropionic acid model for convulsion testing

[0130]

[0131] 4. Quantitative pharmacological evaluation experiments.

[0132] Pharmacological quantitative evaluation experiments involve calculating the median effective dose (LD50) and median neurotoxicity of the anticonvulsant activity, and simultaneously calculating the corresponding 95% confidence limits; the median effective dose is defined as the ED50. 50 The median neurotoxicity dose was TD. 50 .

[0133] Animals were randomly divided into groups of 8. As shown in Table 6, each group received a dose between the lowest and highest doses. The lowest dose group was defined as a dose group in which less than or equal to 20% of mice showed a positive result. The highest dose group was defined as a dose group in which more than or equal to 80% of mice showed a positive result. The interval between the lowest and highest dose groups was divided according to a geometric progression, typically with a ratio of 1:1.2, with each value representing one dose group, containing 3 to 4 dose groups. The calculation was performed using the following formula:

[0134] ED 50 =lg -1 [X m -i(∑P-0.5)];

[0135] SX 50 =i(∑P-∑P 2 / n-1) 1 / 2 ;

[0136] ED 50 95% confidence limit = lg -1 (lgED 50 ±1.96SX 50 );

[0137] In the formula, X m The common logarithm of the maximum dose; i represents the common logarithm of the ratio between doses; P represents the positive rate of each group, expressed as a decimal; n represents the number of animals in each group; SX 50 lgED 50 The standard error.

[0138] TD 50 Calculate according to the following formula:

[0139] TD 50 =lg -1 [X m -i(∑P-0.5)];

[0140] SX 50=i(∑P-∑P 2 / n-1) 1 / 2 ;

[0141] TD 50 95% confidence limit = lg -1 (lgTD 50 ±1.96SX 50 );

[0142] In the formula, X m The common logarithm of the maximum dose; i represents the common logarithm of the ratio between doses; P represents the positive rate of each group, expressed as a decimal; n represents the number of animals in each group; SX 50 For lgTD 50 The standard error.

[0143] Table 6 Results of Quantitative Pharmacological Evaluation Experiments

[0144]

[0145] 5. Neurotoxicity evaluation.

[0146] Neurotoxicity was evaluated using a rotating bar test. Mice were placed on a 1-inch diameter corrugated cylinder rotating at 6 rpm 30 min after intraperitoneal administration or 1 h after oral administration. A positive neurotoxicity test was achieved if the mouse could maintain its balance for 1 minute in each of three consecutive tests without falling off the cylinder; otherwise, a negative test was considered ineffective. The TD (digestive toxicity) was calculated using the Bliss method. 50 The protection index (PI) is calculated using the following formula: PI = TD 50 / ED 50 .

[0147] Table 7 Experimental results of neurotoxicity

[0148]

[0149] Pharmacodynamic experiments showed that:

[0150] As shown in Table 2, the half-maximal effective dose (ED) of compound H-10 prepared in Example 1 in the MES model is... 50 The value is 31 mg / kg.

[0151] As shown in Table 5, compound H-10 exhibited significant anticonvulsant activity in the 3-MP model, confirming its agonistic effect on the GABAergic system. Compared to the positive control drug carbamazepine, compound H-10 effectively controlled 3-MP-induced epilepsy.

[0152] As shown in Table 6, it exhibits excellent activity and extremely high safety in a subcutaneous pentylenetetrazol-induced epilepsy model, with an ED50% (expiratory value of 6.5%). 50As low as 8.86 mg / kg.

[0153] As shown in Table 7, the median neurotoxicity dose (TD) 50 The efficacy was as high as 93.67 mg / kg, with a protection index (PI) of 10.6. This PI value is significantly higher than that of carbamazepine, a first-line clinical drug, under the same model.

[0154] This demonstrates that the compounds synthesized in the embodiments of the present invention have an extremely broad therapeutic window and excellent drug safety for GABAergic system-related epilepsy types.

[0155] Mechanism of action study:

[0156] 1. MOE molecular docking.

[0157] like Figure 2 MOE molecular docking simulations showed that H-10 can stably bind to the classical benzodiazepine site of the GABA_A receptor, with a binding free energy (ΔG) of -8.0603 kcal / mol, indicating a highly spontaneous thermodynamic binding process. The binding mode of H-10 reveals the structural reasons for its high affinity: Key hydrogen bonding: The triazole group of the compound forms a stable hydrogen bond network with Asn265 residues on the B chain and Gln229 residues on the A chain, acting as "anchors" to precisely fix the molecule at the entrance of the binding pocket. π-π stacking: The benzene ring structure in the molecule generates a strong face-to-face π-π stacking interaction with Phe289 residues on the B chain, providing a significant contribution to the binding energy. Hydrophobic cavity encapsulation: The docking model shows that the heptyl side chain of H-10 is fully extended and embeds itself into a narrow tunnel-like cavity composed of hydrophobic amino acid residues by changing its conformation, achieving perfect spatial and hydrophobic complementarity. This provides a structural biological explanation for its high binding affinity and excellent antiepileptic activity.

[0158] 2. Western blotting experiment.

[0159] Western blot analysis was used to quantitatively analyze the expression changes of key proteins in the GABAergic pathway in hippocampal tissue.

[0160] Kunming mice weighing 18g–22g were randomly divided into five groups (n=3 / group): a blank control group, a PTZ model group, a low-dose H-10 group, a medium-dose H-10 group, and a high-dose H-10 group. The blank control group was designated as Control, the PTZ model group as PTZ, and the dose in the low-dose H-10 group as ED. 50 The dose in the H-10 medium-dose group was 2ED. 50 The dose in the H-10 high-dose group was TD. 50Except for the Control group, all other groups received a subcutaneous injection of PTZ at a dose of 100 mg / kg 30 minutes after administration.

[0161] Hippocampal tissue was harvested 30 minutes after modeling, and lysis buffer containing protease inhibitors was added. The tissue was mechanically ground with zirconia beads and lysed in an ice bath for 30 minutes. After centrifugation at 12,000 rpm for 30 minutes at 4°C, the supernatant was collected for protein quantification. Immunoblotting analysis was performed using four proteins: ABAT, GAT1, GABRA1, and CAD65, with GAPDH as an internal control. This experiment aimed to further confirm the mechanism of action of compound H-10.

[0162] like Figure 3 As shown, compound H-10 relieves the brain's ineffective and chaotic compensatory state by enhancing GABA receptor function. Downregulation of ABAT enhances receptor function, increasing GABA signaling efficiency and eliminating the need for extensive GABA degradation to maintain balance. Similarly, small amounts of GABA inhibit the receptors enhanced by compound H-10, reducing the need for extensive GABA degradation to maintain balance. The strong inhibitory signal prevents the synaptic cleft from rushing to reuptake GABA to terminate the signal, leading to decreased GAT1 expression. This compound is a positive allosteric regulator, enhancing GABAergic inhibitory signals. When PTZ weakens the inhibitory strength of GABAergic pathways, compound H-10 enhances receptor function. Neurons do not need internalization for self-preservation; instead, they respond positively to therapeutic signals by increasing receptor expression, further strengthening the inhibitory pathway. This indicates that compound H-10 not only provides an acute effect but also induces long-term adaptive changes, reversing the pathological state caused by the PTZ model.

[0163] This indicates that compound H-10 dose-dependently downregulates the expression of ABAT and GAT1 proteins and upregulates the expression of GABRA1 protein in the hippocampus. This suggests that it synergistically enhances GABAergic inhibitory signaling through multiple mechanisms, including reducing GABA degradation, inhibiting reuptake, and increasing receptor expression.

[0164] 3. Oral bioactivity and pharmacokinetics.

[0165] Oral activity test: Mice were randomly divided into 6 groups (n=8 / group) and administered compound H-10 orally at a dose of 20 mg / kg. Subsequently, PTZ at a dose of 100 mg / kg was injected subcutaneously at 15 min, 30 min, 45 min, 60 min, 90 min, and 120 min after administration. Seizure intensity was graded according to the Racine scale (≥IV grade was defined as tonic seizure), and the complete protection rate (100% no tonic seizure) was used as the criterion for determining peak activity.

[0166] like Figure 4As shown, the oral bioactivity window revealed that the complete protection rate of mice reached 100% at 45 minutes after administration, significantly higher than at other time points. The efficacy-time curve exhibited a typical single-peak distribution, with maximum protective efficacy achieved at 45 minutes, suggesting that compound H-10 reaches peak plasma concentration at this time point after oral administration and crosses the blood-brain barrier to exert a central inhibitory effect.

[0167] The oral pharmacokinetic study used LC-MS / MS to detect the content of H-10 in the blood at different time points. As shown in Table 8, the results indicate that compound H-10 reaches its peak concentration in plasma in a very short time, is an immediate-release absorber, and has high oral bioavailability.

[0168] Table 8. Results of oral pharmacokinetic experiments

[0169]

[0170] Note: t 1 / 2 Half-life: The time required for a drug concentration in the body to decrease by half, measured in hours, reflecting the rate at which the drug is eliminated from the body. max This indicates the time to peak concentration, which is the time it takes for a drug to reach its maximum blood concentration after oral administration, expressed in hours, reflecting the rate of drug absorption. C max Peak concentration (AUC) refers to the highest plasma concentration of a drug reached in the body, expressed in ng / mL, reflecting the extent of drug absorption. AUC 0-t represents the area under the plasma concentration-time curve from the time of administration (0) to the last measurable plasma concentration (t), expressed in h × ng / mL, reflecting the total drug exposure during this period. AUC 0-∞ represents the area under the plasma concentration-time curve from the time of administration (0) to the time of complete elimination from the body (∞), expressed in h × ng / mL, reflecting the overall drug exposure. AUC Extrapolated represents the percentage of extrapolated AUC relative to AUC 0-∞, used to assess the reasonableness of extrapolating the final plasma concentration; a lower percentage indicates a smaller extrapolation effect. MRT 0-α represents the mean residence time, expressed in hours, indicating the average time drug molecules remain in the body, reflecting the drug's retention characteristics.

[0171] 4. Evaluation of chronic epilepsy models.

[0172] Establishment of the pre-ignition model:

[0173] To evaluate the therapeutic potential of compound H-10 in chronic refractory epilepsy, a PTZ ignition model was used to simulate disease progression, with PTZ administered intraperitoneally at a dose of 45 mg / kg every other day. Mice were randomly divided into three groups (n=10 / group): a model control group, a low-dose H-10 group, and a high-dose H-10 group. The specific administration regimen was as follows: the low-dose and high-dose H-10 groups received an intraperitoneal injection of compound H-10, followed by PTZ 30 minutes later. All solutions were prepared with sterile deionized water, and PTZ was prepared and used immediately. Specifically, the low-dose H-10 group received a dose of 2 mg / kg; the high-dose H-10 group received a dose of 4.5 mg / kg; and the model control group received an intraperitoneal injection of DMSO, followed by PTZ 30 minutes later.

[0174] The chronic ignition model was established, as shown in Tables 9 and 10:

[0175] Table 9 Experimental arrangement of the chronic ignition model

[0176]

[0177] Table 10 Grouping arrangement of the chronic ignition model

[0178]

[0179] The behavioral scoring criteria for epilepsy are as follows:

[0180] 0 points: No seizure; 1 point: Facial twitching; 2 points: Head nodding / tail raising; 3 points: Forelimb clonus; 4 points: Generalized tonic-clonic jerks.

[0181] Record parameters:

[0182] Incubation period: the time from PTZ injection to the first appearance of symptoms ≥3 points; Duration: the time from the first occurrence of 3 minutes to recovery to <3 minutes; Maximum attack intensity: only the peak value of a single test is recorded.

[0183] like Figure 5 As shown, by using low-dose prophylaxis for refractory epilepsy, compound H-10 at a high dose of 4.5 mg / kg significantly delayed seizures, with a latency period prolonged by 13.8 days compared to the PTZ group; this indicates that compound H-10 effectively raises the seizure threshold. It also simultaneously shortens seizure duration, reducing it by 80%, demonstrating its efficacy in inhibiting epileptic spread. Furthermore, it maximally reduced seizure severity, decreasing the maximum intensity score from 3 (the duration of epileptic clonic episode) to 1.2 (limited to the rhythmic convulsion-tail-erector transition period).

[0184] like Figure 6As shown, in the post-ignition experiment of the chronic ignition model, compound H-10 can still reduce the ignition score and the duration of epileptic seizures. Compared with the positive control drug sodium valproate (300 mg / kg), it can achieve the same protective rate as the positive control drug at medium doses, and is even better than the positive control drug at high doses. In the post-treatment and post-recovery experiments, the compound can treat the epileptic damage caused by PTZ, reduce the severity of seizures, and shorten the duration of seizures.

[0185] This indicates that compound H-10 exhibits significant therapeutic effects in both the pre-ignition and chronic ignition models, confirming its potential for treating refractory epilepsy.

[0186] 5. HE staining.

[0187] like Figures 7-9 Histopathological results showed that the epilepsy model successfully induced specific damage to the brain, heart, and liver. However, mice treated with effective doses of compound H-10 did not develop significant neurotoxic damage after long-term administration. On the contrary, compound H-10 could intervene in and effectively alleviate the pathological changes in these tissues, demonstrating clear neuroprotective and organ-protective effects, and this protective effect was dose-dependent.

[0188] Pharmacodynamic experiments showed that the compounds synthesized in the embodiments of this invention possess potent anticonvulsant activity, particularly exhibiting high activity and high safety in the scPTZ model. Mechanism-of-action studies suggest its potential as a positive allosteric regulator of GABA_A receptors, capable of inducing long-term adaptive changes. It demonstrated therapeutic efficacy in various acute and chronic epilepsy models, and also exhibited neuroprotective and organ-protective effects.

[0189] It should be noted that when numerical ranges are involved in this invention, it should be understood that the two endpoints of each numerical range, as well as any value between the two endpoints, can be selected. Since the steps and methods used are the same as in the embodiments, preferred embodiments are described in this invention to avoid redundancy. Although preferred embodiments of this invention have been described, those skilled in the art, once they understand the basic inventive concept, can make other changes and modifications to these embodiments, and all such changes and modifications fall within the scope of this invention.

[0190] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. If these modifications and variations fall within the scope of equivalents of this invention, then this invention also intends to include these modifications and variations.

Claims

1. An arylalkylazole derivative, characterized in that, The chemical structural formula of the arylalkyl azole derivative is as follows: ； wherein R is a C1-C 10 alkyl group.

2. The arylalkyl azole derivative according to claim 1, characterized in that, R is -C7H 15 .

3. A process for the preparation of the arylalkyl oxazole derivative according to claim 1 or 2, characterized in that, The method comprises the following steps: The intermediate 1 is subjected to a cyclization reaction with (E)-N'-[(E)-(dimethylamino)methylene]-N,N-dimethylhydrazinecarboxamide under the action of a catalyst to obtain an intermediate 2; the catalyst is p-toluenesulfonic acid; The intermediate 2 is subjected to a demethylation reaction with boron tribromide to obtain an intermediate 3; Under alkaline conditions, the intermediate 3 is subjected to a nucleophilic substitution reaction with a haloalkane in a solvent to obtain the arylalkyl azole derivative; The specific synthesis route is as follows: 。 4. The method of producing an arylalkyl azole derivative according to claim 3, wherein The molar ratio of the intermediate 1 to (E)-N'-[(E)-(dimethylamino)methylene]-N,N-dimethylhydrazinecarboxamide is 1:1-2; The molar ratio of the intermediate 2 to boron tribromide is 1:2; The molar ratio of the intermediate 3 to the haloalkane is 1:3-5.

5. The method for preparing arylalkylazole derivatives according to claim 3, characterized in that, The haloalkane is n-heptyl bromide, ethyl bromide, propyl bromide, butyl bromide, isopentyl bromide, 2-methylbutyl bromide, 4-methylpentyl bromide, 5-methylhexyl bromide, n-hexyl bromide, n-octyl bromide or n-decyl bromide.

6. The method for preparing arylalkylazole derivatives according to claim 3, characterized in that, The solvent is at least one of acetonitrile, acetone and N,N-dimethylformamide; and the base is potassium carbonate.

7. The method for preparing arylalkylazole derivatives according to claim 3, characterized in that, The specific preparation method of the intermediate 2 is as follows: After the intermediate 1 is mixed with (E)-N'-[(E)-(dimethylamino)methylene]-N,N-dimethylhydrazinecarboxamide, a catalyst is added under ice bath, and a cyclization reaction is performed at 135-145 DEG C to obtain the intermediate 2.

8. The method for preparing arylalkylazole derivatives according to claim 3, characterized in that, The specific preparation method of the intermediate 3 is as follows: After the intermediate 2 is mixed with a solvent, a boron tribromide solution is added at 0 DEG C, and a demethylation reaction is performed at room temperature to obtain the intermediate 3; the solvent is dichloromethane.

9. Use of an arylalkyl azole derivative for the manufacture of a medicament for the prophylaxis or treatment of chronic nervous system diseases, characterized in that, The arylalkyl azole derivative is the arylalkyl azole derivative according to claim 1 or 2.

10. Use of the arylalkyl azole derivative according to claim 9 for the manufacture of a medicament for the prophylaxis and treatment of chronic nervous system diseases, characterized in that, The chronic nervous system disease is an epilepsy disease.

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