Benzoxa(thia)azepine compounds and their medical use

Abstract

This invention discloses a benzo[a]oxo(carbon / sulfur)nitrogen compound and its applications. The general structural formulas of the benzo[a]oxo(carbon / sulfur)nitrogen compounds of this invention are shown in formulas (I) and (II): wherein: A is selected from methyl (racemic, R configuration, S configuration), ethyl (racemic, R configuration, S configuration), or cyclopropyl; X1 is selected from oxygen, ammonia, or sulfur; X2 is selected from methylene, oxygen, or sulfur; R1 is selected from hydrogen, fluorine, chlorine, bromine, or methyl; R2 is selected from hydrogen, fluorine, chlorine, bromine, or methyl. The benzo[a]oxo(carbon / sulfur)nitrogen compounds of this invention exhibit good monoamine oxidase B inhibitory activity and show extremely high selectivity within the monoamine oxidase family; in a mouse Parkinson's disease model, the benzo[a]oxo(carbon / sulfur)nitrogen compounds of this invention show good therapeutic effects.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to a benzo[a]oxo(carbon / sulfur)nitrogen heterocyclic compound and its applications. Background Technology

[0002] Neurodegenerative diseases have long been considered one of the most mysterious and challenging problems in the biomedical field. As research on neurodegenerative diseases has shifted from descriptive phenomenology to mechanistic analysis, it has become increasingly clear that their pathogenesis is often caused by a combination of multiple factors (genetic, environmental, and endogenous). Common mechanisms include protein misfolding and aggregation, oxidative stress and free radical formation, metal homeostasis dysregulation, mitochondrial dysfunction, and protein phosphorylation.

[0003] Parkinson's disease (PD) is the second most serious neurodegenerative disease after Alzheimer's disease (AD). Studies have shown that monoamine oxidase B (MAO-B) is involved in the development and progression of Parkinson's disease. Inhibiting MAO-B activity can increase dopamine (DA) levels, thereby improving symptoms in PD patients. DA loss is considered a pathological marker of Parkinson's disease, and current treatment strategies primarily focus on increasing DA levels in the brain. Various drugs to alleviate Parkinson's symptoms have been developed and are used clinically, among which MAO-B inhibitors have shown better efficacy and a more favorable safety profile.

[0004] Currently, clinically used MAO-B inhibitors can be divided into three generations: Selegiline is a first-generation irreversible MAO-B inhibitor. Its chemical structure belongs to phenylethylamine derivatives. After metabolism in the body, the metabolites are amphetamine derivatives with sympathomimetic activity. These sympathomimetic metabolites increase the risk of heart disease and hypertension in patients taking selegiline to treat Parkinson's disease (PD). Due to its poor selectivity and significant adverse reactions, it is not a first-line drug for PD treatment. Rasagiline is a second-generation irreversible selective MAO-B inhibitor. Its pharmacodynamic group is propargylamine, which makes rasagiline more effective at irreversibly inhibiting monoamine oxidase, thus exerting a better neuroprotective effect. Furthermore, its metabolite is an inactive non-phenylethylamine substance with fewer toxic side effects, making it a commonly used MAO-B inhibitor for PD treatment in clinical practice. However, because rasagiline has poor selectivity for MAO-B and its inhibition of enzyme activity is irreversible, it is still not the ideal drug for treating PD. The third-generation MAO-B inhibitor is safinamide, a reversible and selective MAO-B inhibitor. Unlike traditional MAO-B inhibitors such as selegiline and rasagiline, safinamide has higher selectivity for MAO-B and its effects are reversible, making it safer for clinical use. Safinamide also inhibits the release of glutamate and the reuptake of dopamine and serotonin. However, some patients may experience discomfort such as difficulty moving, decreased sleep quality, or nausea after taking this drug. Currently, the variety of selective MAO-B inhibitors available in clinical use is very limited, making it difficult to meet the diverse treatment needs. Therefore, the research and development of reversible, highly selective monoamine oxidase B inhibitors, i.e., third-generation monoamine oxidase inhibitors, has become a hot topic in this field. Summary of the Invention

[0005] To address the shortcomings of the prior art, this invention provides a benzo[a]oxo(carbon / sulfur)nitrogenous compound and its application in medicaments for the prevention and / or treatment of diseases related to monoamine oxidase B. The benzo[a]oxo(carbon / sulfur)nitrogenous compound of this invention exhibits excellent monoamine oxidase B inhibitory activity and demonstrates extremely high selectivity within the monoamine oxidase family.

[0006] The benzo[a]oxo(carbon / sulfur)nitrogen heterocyclic compounds of the present invention have the following general structural formulas (I) and (II):

[0007]

[0008] in:

[0009] A is selected from methyl (racemic, R configuration, S configuration), ethyl (racemic, R configuration, S configuration), or cyclopropyl; X1 is selected from oxygen, ammonia, or sulfur; X2 is selected from methylene, oxygen, or sulfur; R1 is selected from hydrogen, fluorine, chlorine, bromine, or methyl; R2 is selected from hydrogen, fluorine, chlorine, bromine, or methyl.

[0010] Furthermore, the benzo[a]oxo(carbon / sulfur)nitrogen compounds are selected from the following structures:

[0011]

[0012] The present invention relates to the application of benzo[a]oxo(carbon / sulfur)nitrogen compounds in the preparation of pharmaceutical formulations.

[0013] The pharmaceutical preparation is a pharmaceutical preparation for the prevention and / or treatment of diseases related to monoamine oxidase B.

[0014] The diseases associated with monoamine oxidase B include, but are not limited to, neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, malignant tumors, depression, and anxiety.

[0015] The present invention also provides a pharmaceutical composition comprising the benzo[a]oxo(carbon / sulfur)nitrogenous compounds or their pharmaceutically acceptable salts, eutectics or solvates.

[0016] The pharmaceutical composition also includes pharmaceutically acceptable excipients and carriers.

[0017] The pharmaceutical composition is in the form of tablets, capsules, powders, granules, syrups, solutions, oral liquids, tinctures, aerosols, powder inhalers, injections, sterile powders for injection, or suppositories.

[0018] The pharmaceutical composition is administered orally, intravenously, intramuscularly, or subcutaneously.

[0019] Specifically, the benzo[a]oxo(carbon / sulfur)nitrogen compounds can also exist in the drug as their solvates or as pharmaceutically acceptable salts. The term "pharmaceutically acceptable salt" refers to a salt formed by the compound with an acid or base that is suitable for use as a drug, including both inorganic and organic salts. A preferred class of salts in this invention are salts formed by benzo[a]oxo(carbon / sulfur)nitrogen compounds with acids. Suitable acids for salt formation include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid.

[0020] Specifically, the drug also includes a pharmaceutically acceptable carrier. A "pharmaceutically acceptable" component is a substance suitable for use in humans or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., a reasonable benefit / risk ratio. A "pharmaceutically acceptable carrier" is a pharmaceutically or food-grade solvent, suspending agent, or excipient used to deliver the compounds of the present invention to animals or humans. The carrier can be liquid or solid. More specifically, pharmaceutically acceptable carriers include various pharmaceutically commonly used excipients and / or excipients, including but not limited to sugars (such as lactose, glucose, and sucrose), starches (such as corn starch and potato starch), cellulose and its derivatives (such as sodium carboxymethyl cellulose, ethyl cellulose, and methyl cellulose), tragacanth powder, malt, gelatin, talc, solid lubricants (such as stearic acid and magnesium stearate), calcium sulfate, vegetable oils (such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and cocoa butter), polyols (such as propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol), malonic acid, emulsifiers (such as Tween / PVC castor oil), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, isotonic salt solutions, and phosphate buffers, etc.; the carrier can improve the stability, activity, and bioavailability of the formulation as needed.

[0021] Specifically, the drug is in the form of tablets, capsules, powders, granules, syrups, solutions, oral liquids, tinctures, aerosols, powder inhalers, injections, sterile powders for injection, or suppositories.

[0022] The beneficial effects of this invention are as follows: Based on reports that benzyloxyaryl derivatives can effectively inhibit MAO-B, and taking into account the subtle differences between the two targets, MAO-A and MAO-B, particularly the differences between the two receptor amino acid residues, a series of molecular structures expected to have good inhibitory activity and selectivity against MAO-B were designed using a structure-based drug molecule design method. Through chemical synthesis and further bioactivity testing, it was shown that among benzo[a]oxo(carbon / sulfur)nitrogenous compounds, the presence of a seven-membered oxygen-containing ring exhibits excellent activity against MAO-B and high selectivity within the MAO family, confirming the potential of this series of compounds to treat diseases regulated by MAO-B. Further biological experiments confirmed the superiority of some compounds in the treatment of Parkinson's disease. Attached Figure Description

[0023] Figure 1 This is a verification of the reversibility of compounds 17 and 19 of the present invention.

[0024] Figure 2 It is a trajectory diagram of an open field experiment.

[0025] Figure 3These are the results of three behavioral studies conducted on MPTP-induced PD mice. (A) Total distance traveled by the mice; (B) Number of times the mice entered the central region; (C) Time spent in the central region by the mice; (D) Average speed of movement of the mice; (E) Time spent on the stick; (F) Score of the mouse grasping test.

[0026] Note: All medications were administered via intraperitoneal injection. Statistical significance was analyzed using t-tests, and data are expressed as Mean ± SEM (n = 6; ####p < 0.0001, control group vs. MPTP group; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, inhibitor groups vs. MPTP group). Detailed Implementation

[0027] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, 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.

[0028] In all embodiments of the present invention 1 The H NMR spectra were obtained using a 600M superconducting nuclear magnetic resonance spectrometer (AVANCE NEO600), and chemical shifts were expressed in ppm using a tetramethylsilane internal standard (0.00 ppm). 1 H NMR representation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of doublet, dt = doublet of triplet. If the coupling constant is provided, the unit is Hz.

[0029] Thin-layer chromatography silica gel plates are Yantai Huanghai HSGF254 or Qingdao GF254. The silica gel plates used in thin-layer chromatography (TLC) have a diameter of 0.15~0.2 mm, and the diameter of the thin-layer chromatography separation and purification products is 0.4~0.5 mm.

[0030] Column chromatography used Yantai Huanghai silica gel 200~300 mesh as the carrier.

[0031] Example 1:

[0032]

[0033] The synthesis route is as follows:

[0034]

[0035] Step 1: Synthesis of methyl 3-fluorobenzyloxy-4-hydroxybenzoate

[0036] Methyl 2,4-dihydroxybenzoate (6.3 g, 1.2 equiv) was placed in a round-bottom flask. Sodium carbonate (11.95 g, 2.5 equiv) was added to 800-120 mL acetonitrile as solvent, and the mixture was stirred at room temperature for 15 min. Then, p-fluorobenzyl bromide (5 g, 1.2 equiv) was added, and the mixture was stirred for 2 min. The mixture was then heated to 78 °C and refluxed for 4 h. The reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the solvent was evaporated, and the product was extracted and subjected to column chromatography to obtain the target compound (4.25 g, yellow oil, yield 47.2%).

[0037] Step 2: Synthesis of methyl 2-(2-((tert-butoxycarbonyl)amino)ethoxy)-4-((3-fluorobenzyl)oxy)benzoate

[0038] Methyl 3-fluorobenzyloxy-4-hydroxybenzoate (1.73 g, 1 equiv), tert-butyl carbamate (2-hydroxyethyl) (1.21 g, 1.2 equiv), and triphenylphosphine (3.28 g, 2 equiv) were placed in a round-bottom flask with 30–50 mL of tetrahydrofuran as solvent. The mixture was stirred at 0 °C for 5 min, and diisopropyl azodicarbonate (1.77 g, 1.4 equiv) was added dropwise. The mixture was kept under N2 protection and stirred at room temperature for 24 h. The reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the solvent was evaporated, and the product was extracted and subjected to column chromatography to obtain the target compound (1.8 g, white oil, yield 68.7%).

[0039] Step 3: Synthesis of methyl 2-(2-aminoethoxy)-4-(3-fluorobenzyl)oxy)benzoate

[0040] 1.8 g of methyl 2-(2-((tert-butoxycarbonyl)amino)ethoxy)-4-((3-fluorobenzyl)oxy)benzoate (1 equiv) was placed in a round-bottom flask with 30-50 mL of dichloromethane as solvent. The mixture was stirred at 0 °C for 5 min, and 10-20 mL of trifluoroacetic acid was added dropwise. The mixture was stirred at room temperature for 1 h, and the reaction was monitored by thin-layer chromatography (TLC). After the reaction was completed, the mixture was neutralized with saturated sodium carbonate, and the target compound (1.1 g, white oil, yield 80.3%) was obtained by extraction and column chromatography.

[0041] Step 4: Synthesis of 8-[(3-fluorophenyl)methoxy]-2,3,4,5-tetrahydro-1,4-benzoxazolin-5-one

[0042] Methyl 2-(2-aminoethoxy)-4-(3-fluorobenzyl)oxy)benzoate (1.1 g, 1 equiv) was placed in a round-bottom flask, and 10-20 mL of methanol was used as solvent. The mixture was stirred at 0 °C for 5 min, and sodium methoxide (1.4 g, 8 equiv) was added. The mixture was stirred at room temperature for 9 h, and the reaction was monitored by thin-layer chromatography (TLC). After the reaction was completed, the solution was evaporated to dryness, and the target compound (0.7 g, white oil, yield 71.4%) was obtained by extraction and column chromatography.

[0043] Step 5: Synthesis of 8-[(3-fluorophenyl)methoxy]-2,3,4,5-tetrahydro-1,4-benzoxazolino

[0044] 8-[(3-fluorophenyl)methoxy]-2,3,4,5-tetrahydro-1,4-benzoxazolin-5-one (0.7 g, 1 equiv) was placed in a round-bottom flask. Using 10-20 mL of tetrahydrofuran as solvent, aluminum chloride (0.49 g, 1.5 equiv) was added. The mixture was stirred at -20°C for 5 min, followed by the addition of lithium aluminum hydride (0.37 g, 4 equiv). After stirring for 2 min, the mixture was heated to 60°C and refluxed for 6 h. The reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the solution was quenched with 10% sodium hydroxide aqueous solution. The filtrate was evaporated to dryness, and the resulting product was extracted and subjected to column chromatography to obtain the target compound (0.5 g, white oil, yield 66.6%).

[0045] Step 6: Synthesis of 2-{8-[(3-fluorophenyl)methoxy]-2,3,4,5-tetrahydro-1,4-benzoxazolin-4-yl}propionamide

[0046] 8-[(3-fluorophenyl)methoxy]-2,3,4,5-tetrahydro-1,4-benzoxazolinone (100 mg, 1 equiv) was placed in a round-bottom flask. Using 5–10 mL of N,N-dimethylformamide as solvent, sodium carbonate (127 mg, 2.5 equiv) was added, and the mixture was stirred at room temperature for 15 min. Then, 2-aminopropionamide (67.2 mg, 1.2 equiv) and potassium iodide (6 mg, 0.1 equiv) were added, and the mixture was stirred for 2 min. The mixture was then heated to 110 °C and refluxed for 4 h. The reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the target compound (80 mg, white solid, yield 62.9%) was obtained by extraction and column chromatography.

[0047] 1H NMR (600 MHz, DMSO-d6) δ (ppm) 7.43 (td, J = 8.0, 6.0 Hz, 1H), 7.29-7.23 (m, 2H), 7.19-7.13 (m, 2H), 7.08-6.99 (m, 2H), 6.67-6.60 (m, 2H), 5.09 (s,2H), 4.00 (dddd, J = 38.7, 12.6, 6.4, 2.6 Hz, 2H), 3.75-3.57 (m, 2H), 3.21 (q,J = 6.8 Hz, 1H), 3.03-2.89 (m, 2H), 1.25 (d, J = 19.0 Hz, 1H), 1.13 (d, J = 6.8Hz, 3H).

[0048] Example 2:

[0049]

[0050] The preparation steps were the same as in Example 1, except that methyl 2,4-dihydroxybenzoate was replaced with methyl 2,5-dihydroxybenzoate, while other conditions remained unchanged.

[0051] 1 H NMR (600 MHz, Chloroform-d) δ (ppm) 7.37-7.32 (m, 1H), 7.17 (d, J = 7.5Hz, 1H), 7.15-7.11 (m, 1H), 7.03-7.00 (m, 1H), 6.95 (d, J = 8.7 Hz, 1H), 6.81-6.71 (m, 2H), 5.00 (s, 2H), 4.10 (d, J = 7.6 Hz, 1H), 3.99 (d, J = 12.4 Hz, 1H), 3.82-3.59 (m, 2H), 3.40 (s, 1H), 3.09 (d, J = 97.6 Hz, 2H), 1.34 (s, 3H).

[0052] Example 3:

[0053]

[0054] The preparation steps are the same as in Example 1, except that m-fluorobenzyl bromide is replaced with p-fluorobenzyl bromide, and other conditions remain unchanged.

[0055] 1H NMR (600 MHz, Chloroform-d) δ (ppm) 7.42-7.36 (m, 2H), 7.09-7.05 (m, 2H), 7.01 (d, J = 8.2 Hz, 1H), 6.67-6.57 (m, 2H), 4.98 (s, 2H), 4.18-4.01 (m, 2H), 3.78-3.59 (m, 2H), 3.38 (d, J = 8.1 Hz, 1H), 3.15 (t, J = 10.5 Hz, 1H), 3.01 (s, 1H), 1.33 (d, J = 7.0 Hz, 3H).

[0056] Example 4:

[0057]

[0058] The preparation steps were the same as in Example 1, except that m-fluorobenzyl bromide was replaced with p-fluorobenzyl bromide, methyl 2,4-dihydroxybenzoate was replaced with methyl 2,5-dihydroxybenzoate, and other conditions remained unchanged.

[0059] 1 H NMR (600 MHz, Chloroform-d) δ (ppm) 7.39-7.36 (m, 2H), 7.08-7.05 (m, 2H), 6.94 (d, J = 8.6 Hz, 1H), 6.79-6.75 (m, 1H), 6.71 (d, J = 3.1 Hz, 1H), 4.95 (s, 2H), 4.08-3.94 (m, 2H), 3.67 (d, J = 41.3 Hz, 2H), 3.38 (s, 1H), 3.15 (s, 1H), 3.00 (s, 1H), 1.33 (d, J = 7.0 Hz, 3H).

[0060] Example 5:

[0061]

[0062] The synthesis route is as follows:

[0063]

[0064] Step 1: Synthesis of 7-methoxy-2,3,4-5-tetrahydro-1H-benzozazepine-1-one

[0065] 6-Methoxy-3,4-dihydronaphthyl-1(2H)-one (1 g, 1 equiv) was placed in a round-bottom flask, and 10–20 mL of methanesulfonic acid was used as solvent. The mixture was stirred at 0 °C for 5 min, and sodium azide (1.1 g, 3 equiv) was slowly added. The mixture was stirred at room temperature for 18 h, and the reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the solvent was evaporated, and the target compound (0.9 g, white solid, yield 83.3%) was obtained by extraction and column chromatography.

[0066] Step 2: Synthesis of 7-hydroxy-2,3,4-5-tetrahydro-1H-benzozazepine-1-one

[0067] 0.9 g (1 equiv) of 7-hydroxy-2,3,4-5-tetrahydro-1H-benzozacycloheptan-1-one was placed in a three-necked flask under N2 atmosphere. 10–20 mL of dichloromethane was used as solvent, and the mixture was stirred at -78 °C for 5 min. Boron tribromide (4.71 g, 5 equiv) was added dropwise using a syringe. The mixture was then slowly heated to 0 °C and reacted for 4 h. The reaction was monitored by thin-layer chromatography (TLC). After the reaction was complete, the solution was quenched with saturated sodium carbonate aqueous solution. Extraction and column chromatography were performed to obtain the target compound (0.75 g, white solid, yield 90.3%).

[0068] Step 3: Synthesis of 7-((4-fluorobenzyl)oxy)-2,3,4-5-tetrahydro-1H-benzozazepine heptane-1-one

[0069] Referring to Example 1, m-fluorobenzyl bromide was replaced with p-fluorobenzyl bromide, and methyl 3-fluorobenzyloxy-4-hydroxybenzoate was replaced with 7-hydroxy-2,3,4-5-tetrahydro-1H-benzozazepine-1-one, while other conditions remained unchanged.

[0070] Steps 4 and 5: Synthesis of 2-(7-((4-fluorobenzyl)oxy)-1,3,4-5-tetrahydro-2H-benzozaheptan-2-yl)propionamide

[0071] Referring to Example 1, 8-[(3-fluorophenyl)methoxy]-2,3,4,5-tetrahydro-1,4-benzoxazolin-5-one was replaced with 7-((4-fluorobenzyl)oxy)-2,3,4-5-tetrahydro-1H-benzozaheptan-1-one, while other conditions remained unchanged.

[0072] 1H NMR (600 MHz, DMSO-d6) δ (ppm) 7.51-7.45 (m, 2H), 7.25-7.18 (m, 2H), 7.14-6.94 (m, 3H), 6.86-6.80 (m, 1H), 6.75-6.69 (m, 1H), 5.03 (s, 2H), 3.78-3.53 (m, 2H), 3.08 (q, J = 6.8 Hz, 1H), 2.95 (dd, J = 7.0, 3.9 Hz, 2H), 2.87-2.74 (m, 2H), 2.50 (d, J = 2.1 Hz, 2H), 1.74-1.66 (m, 1H), 1.60 (ddt, J = 13.6, 11.0, 5.3Hz, 1H), 1.11 (dt, J = 6.9 Hz, 3H).

[0073] Example 6:

[0074]

[0075] The preparation steps are the same as in Example 5, except that p-fluorobenzyl bromide is replaced with m-fluorobenzyl bromide, and other conditions remain unchanged.

[0076] 1 H NMR (600 MHz, DMSO-d6) δ (ppm) 7.47-7.40 (m, 1H), 7.30-7.23 (m, 2H), 7.18-7.11 (m, 1H), 7.03-6.93 (m, 3H), 6.82 (d, J = 2.7 Hz, 1H), 6.73-6.68 (m, 1H), 5.08 (s, 2H), 3.75-3.58 (m, 2H), 3.08 (q, J = 6.8 Hz, 1H), 2.95 (dd, J =6.9, 3.9 Hz, 2H), 2.84-2.73 (m, 2H), 1.11 (d, J = 6.9 Hz, 3H).

[0077] Example 7:

[0078]

[0079] The preparation steps are the same as in Example 1, except that m-fluorobenzyl bromide is replaced with m-methylbenzyl bromide, and other conditions remain unchanged.

[0080] 1H NMR (600 MHz, Chloroform-d) δ (ppm) 7.25-7.17 (m, 2H), 7.14 (d, J = 7.5Hz, 1H), 7.00 (d, J = 8.3 Hz, 1H), 6.73-6.60 (m, 2H), 4.98 (s, 2H), 4.18-4.11 (m, 1H), 4.08-4.00 (m, 1H), 3.82-3.56 (m, 2H), 3.37 (q, J = 7.0 Hz, 1H), 3.15 (dd, J= 13.0, 7.9 Hz, 1H), 2.99 (dd, J = 13.8, 6.0 Hz, 1H), 1.37–1.24 (m, 3H).

[0081] Example 8:

[0082]

[0083] The preparation steps are the same as in Example 5, except that p-fluorobenzyl bromide is replaced with m-methylbenzyl bromide, and other conditions remain unchanged.

[0084] 1 H NMR (600 MHz, Chloroform-d) δ (ppm) 7.26-7.21 (m, 2H), 7.16-7.12 (m, 1H), 7.01-6.98 (m, 1H), 6.81-6.78 (m, 1H), 6.72-6.68 (m, 1H), 4.99 (s, 2H), 3.65 (q, J = 14.2 Hz, 2H), 3.30 (q, J = 7.0 Hz, 1H), 3.04 (dd, J = 22.6, 13.3 Hz, 2H), 2.91-2.77 (m, 2H), 2.37 (s, 3H), 1.87-1.75(m, 2H), 1.31 (d, J = 7.0 Hz, 3H).

[0085] Example 9:

[0086]

[0087] The preparation steps are the same as in Example 1, except that m-fluorobenzyl bromide is replaced with p-methylbenzyl bromide, and other conditions remain unchanged.

[0088] 1H NMR (600 MHz, Chloroform-d) δ (ppm) 7.33-7.28 (m, 2H), 7.20-7.17 (m, 2H), 7.00-6.98 (m, 1H), 6.67-6.65 (m, 1H), 6.63-6.60 (m, 1H), 5.38 (s, 1H), 4.98 (s, 2H), 4.14 (ddd, J = 12.6, 6.4, 2.1 Hz, 1H), 4.04 (dd, J = 12.9, 7.4 Hz, 1H), 3.72-3.60 (m, 2H), 3.37 (d, J = 7.3 Hz, 1H), 3.14 (t, J = 10.5 Hz, 1H), 3.00 (s, 1H), 1.33 (d, J = 6.9 Hz, 3H).

[0089] Example 10:

[0090]

[0091] The preparation steps are the same as in Example 1, except that m-fluorobenzyl bromide is replaced with benzyl bromide, and other conditions remain unchanged.

[0092] 1 H NMR (600 MHz, Chloroform-d) δ (ppm) 7.37-7.28 (m, 4H), 7.28-7.22 (m, 1H), 7.04-6.89 (m, 2H), 6.61-6.52 (m, 2H), 4.95 (d, J = 4.3 Hz, 3H), 4.09-4.02 (m, 1H), 3.96 (ddd, J = 12.7, 7.5, 2.1 Hz, 1H), 3.67-3.52 (m, 2H), 3.29 (q, J =7.0 Hz, 1H), 3.11-3.02 (m, 1H), 2.96-2.87(m, 1H), 1.25 (d, J = 7.0 Hz, 3H).

[0093] Example 11:

[0094]

[0095] The preparation steps are the same as in Example 5, except that benzyl bromide is replaced with benzyl bromide, and other conditions remain unchanged.

[0096] 1H NMR (600 MHz, Chloroform-d) δ (ppm) 7.45-7.41 (m, 2H), 7.40-7.37 (m, 2H), 7.35-7.31 (m, 1H), 7.04 (s, 1H), 7.02-6.98 (m, 1H), 6.81-6.78 (m, 1H), 6.73-6.68 (m, 1H), 5.55 (d, J = 4.8 Hz, 1H), 5.03 (s, 2H), 3.65 (q, J = 14.3 Hz, 2H), 3.30 (q, J = 7.0 Hz, 1H), 3.11-2.97 (m, 2H), 2.91-2.77 (m, 2H), 1.86-1.74 (m, 1H), 1.30 (d, J = 7.0 Hz, 3H).

[0097] Example 12:

[0098]

[0099] The preparation steps are the same as in Example 5, except that p-fluorobenzyl bromide is replaced with 1-bromo-4-(bromomethyl)-2-fluorobenzene, and other conditions remain unchanged.

[0100] 1 H NMR (600 MHz, Chloroform-d) δ (ppm) 7.57-7.50 (m, 0H), 7.38-7.31 (m, 1H), 7.23-6.98 (m, 4H), 6.83-6.74 (m, 1H), 6.72-6.63 (m, 1H), 5.01 (d, J = 22.7Hz, 2H), 3.65 (q, J = 14.2 Hz, 2H), 3.30 (qd, J = 7.0, 3.2 Hz, 1H), 3.04 (dd, J= 22.0, 12.4 Hz, 2H), 2.90-2.76 (m, 2H), 1.80 (s, 2H), 1.30 (d, J = 7.0 Hz, 3H).

[0101] Example 13:

[0102]

[0103] The preparation steps are the same as in Example 1, except that m-fluorobenzyl bromide is replaced with 1-bromo-4-(bromomethyl)-2-fluorobenzene, and other conditions remain unchanged.

[0104] 1H NMR (600 MHz, Chloroform-d) δ (ppm) 7.37-7.31 (m, 1H), 7.19-7.12 (m, 2H), 7.09-6.98 (m, 3H), 6.67-6.64 (m, 1H), 6.62-6.56 (m, 1H), 5.48-5.40 (m, 1H), 5.02 (s, 2H), 4.17-4.12 (m, 1H), 4.06-4.02 (m, 1H), 3.73-3.62 (m, 2H), 3.37 (q, J= 7.0 Hz, 1H), 3.18-3.12 (m, 1H), 3.00 (dd, J = 13.5, 6.1 Hz, 1H), 1.33 (s, 2H).

[0105] Example 14:

[0106]

[0107] The preparation steps were the same as in Example 1, except that m-fluorobenzyl bromide was replaced with 4-(bromomethyl)-1,2-dichlorobenzene, and other conditions remained unchanged.

[0108] 1 H NMR (600 MHz, Chloroform-d) δ (ppm) 7.57-7.50 (m, 0H), 7.38-7.31 (m, 1H), 7.23-6.98 (m, 4H), 6.84-6.74 (m, 1H), 6.72-6.61 (m, 1H), 5.01 (d, J = 22.7Hz, 2H), 3.65 (q, J = 14.2 Hz, 2H), 3.30 (qd, J = 7.0, 3.2 Hz, 1H), 3.13-2.91 (m, 2H), 2.90-2.73 (m, 2H), 1.80 (s, 2H), 1.30 (d, J = 7.0 Hz, 3H).

[0109] Example 15:

[0110]

[0111] The preparation steps are the same as in Example 1, except that m-fluorobenzyl bromide is replaced with 4-(bromomethyl)-1,2-difluorobenzene, and other conditions remain unchanged.

[0112] 1H NMR (600 MHz, Chloroform-d) δ (ppm) 7.26-7.21 (m, 1H), 7.19-7.08 (m, 2H), 7.03-6.97 (m, 2H), 6.64-6.61 (m, 1H), 6.60-6.56 (m, 1H), 5.94 (d, J = 5.0Hz, 1H), 4.96 (s, 2H), 4.15-4.10 (m, 1H), 4.06-4.00 (m, 1H), 3.71-3.61 (m, 2H), 3.35 (q, J = 7.0 Hz, 1H), 3.16-3.10 (m, 1H), 3.02-2.95 (m, 1H), 1.33 (s, 3H).

[0113] Examples 16-19:

[0114] Chiral separation was performed on Examples 3 and 15 respectively to obtain the products of Examples 16-19.

[0115] The spectral parameters of the products from Examples 16-19 are shown in the table below:

[0116]

[0117] Example 20:

[0118]

[0119] The preparation steps are the same as in Example 5, except that p-fluorobenzyl bromide is replaced with 4-(bromomethyl)-1,2-difluorobenzene, and other conditions remain unchanged.

[0120] 1 H NMR (600 MHz, Chloroform-d) δ (ppm) 7.30-7.27 (m, 1H), 7.21-7.17 (m, 1H), 7.17-7.13 (m, 1H), 7.03 (d, J = 8.2 Hz, 1H), 6.78 (d, J = 2.6 Hz, 1H), 6.68 (dd, J = 8.2, 2.7 Hz, 1H), 5.63 (s, 1H), 5.00 (s, 2H), 3.74-3.63 (m, 2H), 3.35-3.29 (m, 1H), 3.11-3.00 (m, 2H), 2.93-2.86 (m, 1H), 2.85-2.79 (m, 1H), 1.89-1.75 (m, 2H), 1.33 (d, J = 6.9 Hz, 3H).

[0121] Example 21:

[0122]

[0123] The preparation steps are the same as in Example 1, except that m-fluorobenzyl bromide is replaced with m-chlorobenzyl bromide, and other conditions remain unchanged.

[0124] 1 H NMR (600 MHz, Chloroform-d) δ (ppm) 7.41-7.36 (m, 2H), 7.11-7.04 (m, 3H), 7.01 (d, J = 8.2 Hz, 1H), 6.67-6.64 (m, 1H), 6.62-6.59 (m, 1H), 5.58 (s, 1H), 4.98 (s, 2H), 4.15 (ddd, J = 12.8, 6.3, 2.2 Hz, 1H), 4.09-4.02 (m, 1H), 3.69 (q, J = 14.0 Hz, 2H), 3.39 (d, J = 9.3 Hz, 1H), 3.16 (dd, J = 13.5, 7.4 Hz, 1H), 3.02 (dd, J = 14.5, 6.0 Hz, 1H).

[0125] The inhibitory activity of the compounds obtained in the above examples against MAO-A and MAO-B was determined by the following method:

[0126] Dilute MAO-A / B enzymes 800-fold with sodium phosphate buffer (0.05 M, pH=7.4) and set aside. Dissolve approximately 5 mg of the test compound in DMSO solution and dilute with sodium phosphate buffer to the required concentration (ensuring the DMSO content in the prepared solution does not exceed 1%). Add 80 μL of diluted MAO-A or MAO-B enzyme and 20 μL of different concentration gradients of the compound to a black 96-well plate sequentially. For the blank control group, add 20 μL of sodium phosphate buffer. Incubate at 37°C for 15 min to allow the test compound to bind to the MAO-B enzyme, thereby effectively inhibiting the binding of MAO-A / B enzymes to the substrate tyramine. After a 15-minute incubation period, 100 μL of substrate mixture (1100 μL sodium phosphate buffer, 500 μL tyramine, 400 μL horseradish peroxidase, and 20 μL Amplex® Red reagent) was added to each well for reaction, resulting in a total reaction volume of 200 μL. Immediately after substrate addition, fluorescence intensity was measured using a microplate reader (FLX800, Bio-Tek Instruments, Inc., Synergy, HI, USA) (scanning at excitation wavelength of 545 nm and absorption wavelength of 590 nm for 30 minutes at 2-minute intervals). The amount of H₂O₂ produced by the reaction of MAO-B with tyramine was calculated from this data. GraphPad Prism 9.5 software was used to plot curves based on the data collected at each time interval, thereby calculating the IC₂ of the compound. 50 value.

[0127] The results of the bioactivity experiments of compounds 1-21 and positive controls against MAOs are shown in the table below.

[0128]

[0129]

[0130]

[0131]

[0132]

[0133] Long-term use of irreversible MAO-B inhibitors induces compensatory upregulation of diamine oxidase gene expression, resulting in an inability to effectively reduce GABA levels in astrocytes, thus limiting the neuroprotective potential of irreversible MAO-B inhibitors. Therefore, rasagiline, an irreversible inhibitor, was selected as a positive control to evaluate the mechanisms of action of compounds 17 and 19 in relation to hMAO-B.

[0134] The testing method is basically the same as testing hMAO-A and / or hMAO-B ICs. 50 The method is the same; first, a high concentration of inhibitor (50×IC) is used. 50 After incubation for a period of time, the enzyme binds fully to the compound and becomes almost completely inactive. Then, it is rapidly diluted 100-fold with buffer. For reversible inhibitors, the enzyme activity slowly recovers with increasing time after dilution; however, for irreversible inhibitors, the enzyme activity does not recover.

[0135] Figure 1 This verifies the reversibility of compounds 17 and 19. From Figure 1 The test results are consistent with the reported results: rasagiline interacts with hMAO-B through irreversible binding. As the test time increased, the curves of compounds 17 and 19 approached the control group with increasing slope, indicating a decrease in inhibition rate. The enzyme activity gradually recovered with prolonged dilution time, suggesting that compounds 17 and 19 interact with hMAO-B through reversible binding.

[0136] Because patients with Parkinson's disease (PD) typically experience motor dysfunction such as bradykinesia, muscle rigidity, resting tremor, and gait disturbances, their quality of life is severely reduced. Therefore, assessing whether a compound can improve motor dysfunction is an important indicator for pharmacodynamic evaluation. 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a highly lipid-soluble neurotoxin that can rapidly cross the blood-brain barrier and enter the nervous system. Within glial cells, MPTP is metabolized via MAO-B to produce the MPDP+ intermediate, which is further oxidized to form the toxic metabolite MPP+, leading to damage and death of dopaminergic neurons in the SNpc and inducing typical motor symptoms of PD. Based on the excellent MAO-B inhibitory activity of compounds 17 and 19, their pharmacodynamics was investigated in depth in an MPTP-induced subacute PD mouse model.

[0137] C57BL / 6J mice (age: 2 months, weight: 25±2 g) purchased from Henan Scibes Biotechnology Co., Ltd. were randomly divided into the following 7 groups (n = 6 / group): blank group, MPTP group, MPTP + safenamide (30 mg / kg) group, MPTP + compound 17 (10 mg / kg) group, MPTP + compound 17 (30 mg / kg) group, MPTP + compound 19 (10 mg / kg) group, and MPTP + compound 19 (30 mg / kg) group. All groups were administered the medication via intraperitoneal injection. The blank group and MPTP group received an equal volume of solvent intraperitoneally. 30 min later, the MPTP group, compound groups, and safenamide group received an intraperitoneal injection of MPTP solution (20 mg / kg), while the blank group received an equal volume of physiological saline. This procedure was repeated for seven days. On the eighth day, the behavioral performance of the mice was evaluated using the open field test, rotarod test, and grasp test. The specific behavioral experimental methods are as follows:

[0138] (1) The open field test is mainly used to evaluate the spontaneous movement of mice. This test is conducted in darkness, and mice are tested independently after acclimatizing to the dark environment for half an hour. The mice are placed in the open field test apparatus (0.4 × 0.4 × 0.5 m). 3 The mice were placed in a designated area and allowed free movement for 5 minutes. The area was then divided using software, and data such as the mice's movement trajectories and activity times were collected using a camera for analysis.

[0139] (2) The rotundus test was designed to assess the motor abilities of mice. Mice were acclimatized to the instrument and environment one day in advance. During the experiment, mice were placed on a horizontal rotundus, and the instrument was set to uniform acceleration mode, with the rotation speed increasing from 5 rpm to 40 rpm, and then stopped after 3 minutes. The test was conducted from the moment the mouse could remain steadily on the rotundus until it fell off or remained on the rotundus for more than 3 minutes. The time each mouse spent on the rotundus was recorded, and each mouse was tested three times.

[0140] (3) The grasping test mainly assesses the mouse's muscle relaxation performance and motor coordination. The mouse's tail is lifted so that its forepaws grasp a 30 cm long wire 30 cm above the ground. The tail is then released to observe the mouse's grasping ability. The grasping ability is scored according to the following criteria:

[0141] 1: The front paws are hooked onto the metal wire; 2: After the front paws grab the metal wire, try to climb upwards; 3: After grabbing the metal wire with the front paws, one or two hind paws can be used to grab the metal wire; 4: All paws are grabbing the metal wire, and the tail is wrapped around the metal wire; 5: Reach the end of the metal wire and escape from the device.

[0142] Figure 2 It is a trajectory diagram of an open field experiment; Figure 3The results of three behavioral studies on MPTP-induced PD mice are as follows: (A) total distance traveled by the mice; (B) number of times the mice entered the central region; (C) time spent in the central region; (D) average speed of movement; (E) time spent on the stick; and (F) score in the grasp test. Compared with the control group, the MPTP group mice showed significantly reduced time spent on the stick, grasp score, and open field test scores, indicating significant defects in muscle function and motor ability in PD mice. Treatment with safinamide (30 mg / kg) significantly improved all behavioral parameters in the mice. However, compounds 17 or 19 at a dose of 10 mg / kg significantly improved all behavioral indicators in PD model mice, with improvement effects approaching those of the control group. In conclusion, compounds 17 and 19, at low doses, have unique advantages in improving motor dysfunction in MPTP-induced PD mouse models, particularly in improving muscle relaxation and motor coordination.

[0143] In summary, based on the subtle differences between the two targets, MAO-A and MAO-B, particularly the differences between the two receptor amino acid residues, this invention utilizes a structure-based drug design approach to design a series of molecular structures with good inhibitory activity and selectivity against MAO-B. Through chemical synthesis and further bioactivity testing, it was demonstrated that among benzo[a]oxo(carbon / sulfur)nitrogenous compounds, the presence of a seven-membered oxygen-containing ring exhibits excellent activity against MAO-B and high selectivity within the MAO family. In the examples listed in this invention, more than half of the compounds showed superior inhibitory activity and selectivity against MAO-B compared to the positive control drug safenamide. Furthermore, this invention also conducted a jump-diluted assay on two representative compounds, demonstrating that the series of compounds interacts with hMAO-B reversibly. In mouse behavioral studies, compounds 17 and 19 significantly improved MPTP-induced motor dysfunction and muscle damage in mice at low doses. This discovery further confirms the great potential of this series of compounds in treating diseases regulated by MAO-B, providing new ideas and effective drug candidates for the treatment of related diseases, and is expected to play an important role in future clinical applications.

[0144] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A benzo[a]oxo(carbon / sulfur)nitrogen heterocyclic compound, characterized in that... Its general structural formula is shown in equation (Ⅰ): ； in: A is selected from methyl, ethyl, or cyclopropyl; X1 is selected from oxygen or sulfur; X2 is selected from methylene, oxygen, or sulfur; R1 is selected from hydrogen, fluorine, chlorine, bromine, or methyl; R2 is selected from hydrogen, fluorine, chlorine, bromine, or methyl. When A is selected from methyl or ethyl, the compound shown in formula (Ⅰ) is a racemic mixture, R configuration or S configuration.

2. The benzo[a]oxo(carbon / sulfur)nitrogen heterocyclic compound according to claim 1, characterized in that... Compounds selected from the following structures: 。 3. The use of the benzo[a]oxo(carbon / sulfur)nitrogenous compounds of claim 1 in the preparation of monoamine oxidase B inhibitors.

4. The use of the benzo[a]oxo(carbon / sulfur)nitrogen compounds of claim 1 in the preparation of pharmaceutical formulations, characterized in that: The pharmaceutical preparation is a pharmaceutical preparation for the prevention and / or treatment of diseases related to monoamine oxidase B.

5. The application according to claim 4, characterized in that: The diseases associated with monoamine oxidase B are neurodegenerative diseases.

6. The application according to claim 4, characterized in that: The diseases associated with monoamine oxidase B include one or more of the following: Parkinson's disease, Alzheimer's disease, malignant tumors, depression, and anxiety disorders.

7. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the benzoxanox (carbon / sulfur) nitrogen compound of claim 1 or a pharmaceutically acceptable salt thereof.

8. The pharmaceutical composition according to claim 7, characterized in that: The pharmaceutical composition also includes pharmaceutically acceptable excipients.

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