Benzoxy (carbon / sulfur) aza compound and medical application thereof

Abstract

The invention discloses a benzoxo (carbon / sulfur) aza compound and application thereof. The structural general formula of the benzoxo (carbon / sulfur) aza compound is shown as the following formulas (I) and (II): # imgabs0 #, wherein A is selected from methyl (raceme, R configuration and S configuration), ethyl (raceme, R configuration and 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 benzoxo (carbon / sulfur) aza compound disclosed by the invention has good monoamine oxidase B inhibitory activity, and shows extremely high selectivity in a monoamine oxidase family; in a mouse Parkinson's disease model, the benzoxo (carbon / sulfur) aza compound disclosed by the invention shows a relatively good treatment effect.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and specifically relates to a benzooxy (carbon / sulfur) azapropyne compound and application thereof. Background Art

[0002] Neurodegenerative diseases have long been regarded as one of the most mysterious and challenging problems in biomedicine. As the study of neurodegenerative diseases shifts from descriptive phenomenology to mechanistic analysis, it is becoming increasingly clear that their pathogenesis is often caused by multiple factors (genetic, environmental, and endogenous factors). General mechanisms include protein misfolding and aggregation, oxidative stress and free radical formation, metal homeostasis disorders, 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 occurrence and development of Parkinson's disease. Inhibiting the activity of monoamine oxidase with drugs can increase dopamine (DA) levels, thereby improving the symptoms of PD patients. DA loss is considered a pathological hallmark of Parkinson's disease, and current treatment strategies mainly focus on increasing DA levels in the brain. Various drugs that relieve Parkinson's disease symptoms have been developed and used clinically, among which MAO-B inhibitors show better efficacy and more favorable safety.

[0004] The MAO-B inhibitors currently used in clinical practice can be divided into three generations: Selegiline is the first generation of irreversible MAO-B inhibitors. Its chemical structure belongs to phenylethylamine derivatives. After metabolism in the body, the metabolites are amphetamine derivatives with sympathomimetic activity. Metabolites with sympathomimetic activity will increase the risk of heart disease and hypertension in patients taking selegiline to treat PD. Due to its poor selectivity and adverse reactions, it cannot be the first choice for the treatment of PD. The second generation of irreversible MAO-B selective inhibitors is rasagiline. Its pharmacophore is propargylamine, which makes rasagiline more effective in irreversibly inhibiting monoamine oxidase and better exerting neuroprotective effects. Its metabolite is an inactive non-amphetamine substance with little toxicity and side effects. It is currently a commonly used MAO-B inhibitor for the treatment of PD in clinical practice. However, due to the low selectivity of rasagiline for MAO-B and the irreversible inhibition of enzyme activity, it still cannot be the most ideal drug for the treatment of PD. The third generation of MAO-B inhibitors is the reversible MAO-B selective inhibitor safinamide. Unlike traditional MAO-B inhibitors selegiline and rasagiline, safinamide has higher selectivity for MAO-B and its action is 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 symptoms such as difficulty in movement, decreased sleep quality, or nausea after taking the drug. Currently, the types of selective MAO-B inhibitors in clinical use are very limited, and it is difficult to meet the diversity of treatment needs. Therefore, the research and development of reversible and highly selective monoamine oxidase B inhibitors, namely the third generation of monoamine oxidase inhibitors, has become a hot topic in this field. Summary of the invention

[0005] In view of the above-mentioned deficiencies of the prior art, the present invention provides a benzooxy (carbon / sulfur) azepine compound and its use in a drug for preventing and / or treating diseases related to monoamine oxidase B. The benzooxy (carbon / sulfur) azepine compound of the present invention has good monoamine oxidase B inhibitory activity and exhibits ultra-high selectivity within the monoamine oxidase family.

[0006] The benzoxo(carbon / sulfur)azepine compounds of the present invention have the following general structural formulas (I) and (II):

[0007]

[0008] in:

[0009] A is selected from methyl (racemate, R configuration, S configuration), ethyl (racemate, 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] Further, the benzooxy(carbon / sulfur)azepine compound is selected from the following structures:

[0011]

[0012] The invention discloses an application of the benzooxy(carbon / sulfur)azepine compound in the preparation of pharmaceutical preparations.

[0013] The pharmaceutical preparation is a pharmaceutical preparation for preventing and / or treating diseases related to monoamine oxidase B.

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

[0015] The present invention also provides a pharmaceutical composition, which comprises the benzooxy(carbon / sulfur)azepine compound or a pharmaceutically acceptable salt, cocrystal or solvate thereof.

[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, elixirs, aerosols, powder sprays, injections, sterile powders for injection, or suppositories.

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

[0019] Specifically, the benzo(carbon / sulfur)azepine compound can also be present in the drug in the form of its solvate or its pharmaceutically acceptable salt. The term "pharmaceutically acceptable salt" refers to a salt formed by a compound with an acid or a base that is suitable for use as a drug, including inorganic salts and organic salts. A preferred salt of the present invention is a salt formed by a benzo(carbon / sulfur)azepine compound with an acid. Acids suitable for forming salts 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 that is suitable for human beings and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., has a reasonable benefit / risk ratio. A "pharmaceutically acceptable carrier" is a pharmaceutically or food-acceptable solvent, suspending agent, or excipient for delivering the compound of the present invention to an animal or human. The carrier may be a liquid or a solid. More specifically, pharmaceutically acceptable carriers are various excipients and / or vehicles commonly used in pharmacy, 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, glycerol, sorbitol, mannitol and polyethylene glycol), begonia acid, emulsifiers (such as Tween / polyvinyl chloride castor oil), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, isotonic saline solution and phosphate buffer, etc.; the carrier can improve the stability, activity and biological effectiveness of the formulation as needed.

[0021] Specifically, the medicine is a tablet, capsule, powder, granule, syrup, solution, oral liquid, spirit, aerosol, powder spray, injection, sterile powder for injection, or suppository.

[0022] The beneficial effects of the present invention are as follows: Based on the relevant reports that benzyloxyaryl derivatives can effectively inhibit MAO-B, the present invention uses a structure-based drug molecule design method to design a series of molecular structures that are expected to have good inhibitory activity and selectivity against MAO-B, based on the subtle differences between the two targets of MAO-A and MAO-B, especially the differences between the amino acid residues of the two receptors. Through chemical synthesis and further biological activity tests, it is shown that among benzooxy (carbon / sulfur) aza-pyridine compounds, when there is a seven-membered oxygen-containing ring, they will show very excellent activity against MAO-B, and show high selectivity within the MAO family, confirming that this series of compounds has the potential to treat diseases regulated by MAO-B. Further biological experiments confirmed the superiority of some compounds in the treatment of Parkinson's disease. BRIEF DESCRIPTION OF THE DRAWINGS

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

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

[0025] Figure 3The results of three behavioral studies on MPTP-induced PD mice. (A) The total distance moved by the mice; (B) The number of times the mice entered the central area; (C) The time the mice stayed in the central area; (D) The average speed of the mice; (E) The time they stayed on the rod; (F) The score of the mouse grab test.

[0026] Note: All drugs were administered by intraperitoneal injection. Statistical significance was analyzed by t-test, and data were 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 DESCRIPTION

[0027] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0028] In all embodiments of the present invention 1 The H NMR spectra were measured using a 600M superconducting nuclear magnetic resonance spectrometer (AVANCE NEO600). The chemical shifts are expressed in ppm using tetramethylsilane as the internal standard (0.00 ppm). 1 H NMR notation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of a doublet, dt = doublet of a triplet. Coupling constants, when given, are given in Hz.

[0029] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) adopts a specification of 0.15~0.2 mm, and the specification used for thin layer chromatography separation and purification products is 0.4~0.5 mm.

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

[0031] Embodiment 1:

[0032]

[0033] The synthetic route is as follows:

[0034]

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

[0036] Place methyl 2,4-dihydroxybenzoate (6.3 g, 1.2 equiv) in a round-bottom flask, use 800-120 mL of acetonitrile as solvent, add sodium carbonate (11.95 g, 2.5 equiv) and stir at room temperature for 15 min, add p-fluorobenzyl bromide (5 g, 1.2 equiv) and stir for 2 min, then heat to 78°C and reflux for 4 h, and monitor the reaction by TLC. After the reaction is completed, the solvent is dried, and the target compound (4.25 g, yellow oil, yield 47.2%) is obtained by extraction and column chromatography.

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

[0038] Methyl 3-fluorobenzyloxy-4-hydroxybenzoate (1.73 g, 1 equiv), tert-butyl (2-hydroxyethyl) carbamate (1.21 g, 1.2 equiv), and triphenylphosphine (3.28 g, 2 equiv) were placed in a round-bottom flask, and 30-50 mL of tetrahydrofuran was used as solvent. The mixture was stirred at 0°C for 5 min, and diisopropyl azodicarboxylate (1.77 g, 1.4 equiv) was added dropwise. The mixture was protected by N2 and stirred at room temperature for 24 h. The reaction was monitored by TLC. After the reaction was completed, the solvent was dried and the target compound (1.8 g, white oil, yield 68.7%) was obtained by extraction and column chromatography.

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

[0040] Place methyl 2-(2-((tert-butyloxycarbonyl)amino)ethoxy)-4-((3-fluorobenzyl)oxy)benzoate (1.8 g, 1 equiv) in a round-bottom flask, use 30-50 mL of dichloromethane as solvent, stir at 0°C for 5 min, add 10-20 mL of trifluoroacetic acid dropwise, stir at room temperature for 1 h, and monitor the reaction by thin layer chromatography (TLC). After the reaction is completed, neutralize with saturated sodium carbonate, and obtain the target compound (1.1 g, white oil, yield 80.3%) through extraction and column chromatography.

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

[0042] Place methyl 2-(2-aminoethoxy)-4-(3-fluorobenzyl)oxy)benzoate (1.1 g, 1 equiv) in a round-bottom flask, use 10-20 mL of methanol as solvent, stir at 0°C for 5 min, add sodium methoxide (1.4 g, 8 equiv), stir at room temperature for 9 h, and monitor the reaction by TLC. After the reaction is completed, spin dry, extract, and column chromatography to obtain the target compound (0.7 g, white oil, yield 71.4%).

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

[0044] 8-[(3-fluorophenyl)methoxy]-2,3,4,5-tetrahydro-1,4-benzoxazoline-5-one (0.7 g, 1 equiv) was placed in a round-bottom flask, and 10-20 mL of tetrahydrofuran was used as solvent. Aluminum chloride (0.49 g, 1.5 equiv) was added, and stirred at -20°C for 5 min. Lithium aluminum tetrahydride (0.37 g, 4 equiv) was added, and stirred for 2 min, and then the temperature was raised to 60°C and refluxed for 6 h. The reaction was monitored by thin layer chromatography (TLC). After the reaction was completed, the reaction was quenched with 10% sodium hydroxide aqueous solution, and the filtrate was filtered and dried. The target compound (0.5 g, white oil, yield 66.6%) was obtained after extraction and column chromatography.

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

[0046] 8-[(3-fluorophenyl)methoxy]-2,3,4,5-tetrahydro-1,4-benzoxazoline (100 mg, 1 equiv) was placed in a round-bottom flask, 5-10 mL of N,N-dimethylformamide was used as solvent, sodium carbonate (127 mg, 2.5 equiv) was added, and the mixture was stirred at room temperature for 15 min, 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, and then the mixture was heated to 110°C and refluxed for 4 h. The reaction was monitored by thin layer chromatography (TLC). After the reaction was completed, 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] Embodiment 2:

[0049]

[0050] The preparation steps were the same as in Example 1, except that 2,4-dihydroxybenzoic acid methyl ester was replaced with 2,5-dihydroxybenzoic acid methyl ester, and 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] Embodiment 3:

[0053]

[0054] The preparation steps are as in Example 1, except that m-fluorobenzyl bromide is replaced by 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] Embodiment 4:

[0057]

[0058] The preparation steps are as in Example 1, except that m-fluorobenzyl bromide is replaced by p-fluorobenzyl bromide, methyl 2,4-dihydroxybenzoate is replaced by methyl 2,5-dihydroxybenzoate, and other conditions remain 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] Embodiment 5:

[0061]

[0062] The synthetic route is as follows:

[0063]

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

[0065] 6-Methoxy-3,4-dihydronaphthalene-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, the solvent was dried, 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-benzazepan-1-one

[0067] 7-Hydroxy-2,3,4-5-tetrahydro-1H-benzazepan-1-one (0.9 g, 1 equiv) was placed in a three-necked flask under N2 atmosphere, and 10-20 mL of dichloromethane was used as solvent. Stir at -78°C for 5 min, and boron tribromide (4.71 g, 5 equiv) was added dropwise with a syringe. The temperature was slowly raised to 0°C for 4 h, and the reaction was monitored by thin layer chromatography (TLC). After the reaction was completed, the reaction was quenched with saturated sodium carbonate aqueous solvent, and the target compound (0.75 g, white solid, yield 90.3%) was obtained after extraction and column chromatography.

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

[0069] Referring to Example 1, m-fluorobenzyl bromide was replaced by p-fluorobenzyl bromide, 3-fluorobenzyloxy-4-hydroxybenzoic acid methyl ester was replaced by 7-hydroxy-2,3,4-5-tetrahydro-1H-benzazepan-1-one, and other conditions remained unchanged.

[0070] Step 4 to Step 5: Synthesis of 2-(7-((4-fluorobenzyl)oxy)-1,3,4-5-tetrahydro-2H-benzazepan-2-yl)propanamide

[0071] Referring to Example 1, 8-[(3-fluorophenyl)methoxy]-2,3,4,5-tetrahydro-1,4-benzoxazoline-5-one was replaced with 7-((4-fluorobenzyl)oxy)-2,3,4-5-tetrahydro-1H-benzazepan-1-one, and 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] Embodiment 6:

[0074]

[0075] The preparation steps are 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] Embodiment 7:

[0078]

[0079] The preparation steps are 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] Embodiment 8:

[0082]

[0083] The preparation steps are 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] Embodiment 9:

[0086]

[0087] The preparation steps are 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] Embodiment 10:

[0090]

[0091] The preparation steps are 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] Embodiment 11:

[0094]

[0095] The preparation steps are as in Example 5, except that p-fluorobenzyl 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] Embodiment 12:

[0098]

[0099] The preparation steps are 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] Embodiment 13:

[0102]

[0103] The preparation steps are 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] Embodiment 14:

[0106]

[0107] The preparation steps are as in Example 1, except that m-fluorobenzyl bromide is replaced with 4-(bromomethyl)-1,2-dichlorobenzene, and other conditions remain 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] Embodiment 15:

[0110]

[0111] The preparation steps are 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] Embodiments 16 to 19:

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

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

[0116]

[0117] Embodiment 20:

[0118]

[0119] The preparation steps were similar to those of Example 5, except that p-fluorobenzyl bromide was replaced with 4-(bromomethyl)-1,2-difluorobenzene, and other conditions remained 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] Embodiment 21:

[0122]

[0123] The preparation steps are 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 as follows:

[0126] Dilute the MAO-A / B enzyme 800 times with sodium phosphate buffer (0.05 M, pH=7.4) for later use. Then dissolve the weighed (about 5 mg) test compound in DMSO solution and dilute to the required concentration with sodium phosphate buffer (control the DMSO content in the prepared solution to not exceed 1%). Add 80 μL of diluted MAO-A or MAO-B enzyme and 20 μL of compounds with different concentration gradients to the black 96-well plate in sequence. Add 20 μL of sodium phosphate buffer to the blank group, and then incubate in a 37°C incubator for 15 mins to allow the test compound to bind to the MAO-B enzyme, thereby effectively inhibiting the binding of the MAO-A / B enzyme to the substrate tyramine. After a 15-min incubation period, 100 μL of substrate mixed solution (1100 μL sodium phosphate buffer, 500 μL tyramide, 400 μL horseradish peroxidase, 20 μL Amplex® Red reagent) was added to each well for reaction, and the total volume of the entire reaction system was 200 μL. After the addition of the substrate, the fluorescence intensity was immediately detected in a microplate reader (FLX800, Bio-Tek Instruments, Inc., Synergy, HI, USA) (scanning at an excitation wavelength of 545 nm and an absorption wavelength of 590 nm for 30 min, with a time interval of 2 min), and the amount of H2O2 produced by the reaction of MAO-B with tyramide was calculated, and the data read in each time period were fitted with a curve using GraphPad Prism 9.5 software to calculate the IC value of the compound. 50 value.

[0127] The results of the bioactivity experiments of compounds 1 to 21 and the positive control against MAOs are shown in the following table.

[0128]

[0129]

[0130]

[0131]

[0132]

[0133] Long-term use of irreversible MAO-B inhibitors will induce upregulation of compensatory gene expression of diamine oxidase, resulting in the inability to effectively reduce GABA levels in astrocytes, thereby limiting the potential application of irreversible MAO-B inhibitors in neuroprotection. Based on this, the irreversible inhibitor rasagiline was selected as a positive control to evaluate the mode of action of compound 17, compound 19 and hMAO-B.

[0134] Test method basic to testing hMAO-A and / or hMAO-B IC 50 The method is the same, first use a high concentration of inhibitor (50×IC 50 ) Incubate for a period of time, the enzyme fully binds to the compound and is almost completely inactivated, and then quickly dilute it 100 times with buffer. For reversible inhibitors, the activity of the enzyme slowly recovers as the time after dilution increases; for irreversible inhibitors, the activity of the enzyme does not recover.

[0135] Figure 1 This is the reversibility verification of compound 17 and compound 19. Figure 1 It can be seen that the test results are consistent with the reported results: the mode of action of rasagiline and hMAO-B is irreversible binding. As the test time increases, the curves of compound 17 and compound 19 approach the control group and the slope increases, the inhibition rate decreases, and the enzyme activity slowly recovers with the extension of time after dilution, indicating that the mode of action of compound 17 and compound 19 and hMAO-B is reversible binding.

[0136] Since PD patients usually have motor dysfunction such as bradykinesia, muscle rigidity, resting tremor and gait posture disorder, the quality of life of patients is seriously reduced. Therefore, evaluating 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 neurotoxin with good lipid solubility that can quickly cross the blood-brain barrier and enter the nervous system. In glial cells, MPTP is metabolized by MAO-B to generate MPDP+ intermediates, which are further oxidized to form toxic metabolites MPP+, leading to damage and death of dopaminergic neurons in SNpc, inducing typical motor symptoms of PD. Based on the excellent MAO-B inhibitory activity of compounds 17 and 19, their pharmacodynamics were deeply studied in the MPTP-induced subacute PD mouse model.

[0137] C57BL / 6J mice (age: 2 months, weight: 25±2 g) purchased from Henan Sikebes Biotechnology Co., Ltd. were randomly divided into the following 7 groups (n = 6 / group): blank group, MPTP group, MPTP+safinamide (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, MPTP+compound 19 (30 mg / kg) group. All the above groups were administered by intraperitoneal injection, among which the blank group and MPTP group were intraperitoneally injected with equal volumes of solvent. After 30 min, the MPTP group, compound group and safinamide group were intraperitoneally injected with MPTP solution (20 mg / kg), and the blank group was intraperitoneally injected with equal volumes of normal saline. The above operation was repeated for seven days. On the eighth day, the behavioral performance of mice was evaluated by open field test, rotating rod test and snatch test. The specific behavioral experimental methods are as follows:

[0138] (1) The open field test is mainly used to evaluate the autonomous movement of mice. The test is conducted in the dark, and the mice are tested independently after adapting to the dark environment for half an hour. The mice are placed in the open field test device (0.4×0.4×0.5 m 3 The area was divided by software, and the movement trajectory and activity time of the mice were collected by camera for analysis.

[0139] (2) The rotarod test is designed to detect the motor ability of mice's limbs. The mice were allowed to adapt to the instrument and environment one day in advance. During the experiment, the mice were placed on a horizontal rotating rod. The instrument was turned on in uniform acceleration mode, and the rotation speed increased from 5 rpm to 40 rpm and stopped after 3 minutes. The test started from the time when the mouse was able to stay on the rotating rod steadily until it fell off the rotating rod or stayed on the rotating rod for more than 3 minutes. The time each mouse stayed on the rotating rod was recorded, and the experiment was repeated 3 times for each mouse.

[0140] (3) The snatch test mainly evaluates the muscle relaxation performance and movement coordination of mice. The mouse's tail is lifted so that its front paws grasp a metal wire that is 30 cm long and 30 cm above the ground. The tail is then released to observe the mouse's snatch performance. The snatch performance is scored according to the following criteria:

[0141] 1: The front paws hang on the wire; 2: After the front paws grab the wire, try to climb up; 3: After the front paws grab, one or both hind paws can grab the wire; 4: All paws grab the wire and the tail is wrapped around the wire; 5: Reach the end of the wire and escape from the device.

[0142] Figure 2 It is the trajectory diagram of the open field experiment; Figure 3The experimental results of three behavioral studies on MPTP-induced PD mice: (A) total distance moved by mice; (B) number of times mice entered the central area; (C) time spent in the central area of ​​mice; (D) average speed of mouse activity; (E) time spent on the rod; (F) score of the mouse grab test. Compared with the blank group, the time spent on the rod, the grab score and the open field test values ​​of the mice in the MPTP group were significantly reduced, indicating that PD mice had obvious defects in muscle function and motor ability. After treatment with safinamide (30 mg / kg), all behavioral data parameters of mice were significantly improved. However, compound 17 or compound 19 at a dose of 10 mg / kg could significantly improve all behavioral indicators of PD model mice, and the improvement effect was close to the level of the blank group. In summary, compounds 17 and 19 improved motor dysfunction in MPTP-induced PD mouse models at low doses, especially in improving muscle relaxation and improving motor coordination ability.

[0143] In summary, the present invention uses a structure-based drug molecule design method based on the subtle differences between the two targets of MAO-A and MAO-B, especially the differences between the amino acid residues of the two receptors, to design a series of molecular structures with good inhibitory activity and selectivity for MAO-B. Through chemical synthesis and further biological activity tests, it is shown that among benzooxy (carbon / sulfur) aza-pyridine compounds, when there is a seven-membered oxygen-containing ring, they will show very excellent activity against MAO-B and show high selectivity within the MAO family. In the embodiments listed in the present invention, more than half of the compounds are better than the positive control drug safinamide in terms of inhibitory activity and selectivity against MAO-B. In addition, the present invention also conducted a dilution jump experiment on two representative compounds, and the experimental results showed that the mode of action of this series of compounds with hMAO-B is reversible binding. In the mouse behavioral study, compound 17 and compound 19 can significantly improve MPTP-induced mouse motor dysfunction and muscle function damage at low doses. This discovery further confirms that this series of compounds has great potential in treating diseases regulated by MAO-B, provides new ideas and effective drug candidate molecules for the treatment of related diseases, and is expected to play an important role in future clinical applications.

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

Claims

1. A benzooxy(carbon / sulfur)azepine compound, characterized in that Its general structural formula is shown in the following formula (I) and (II): ； in: A is selected from methyl (racemate, R configuration, S configuration), ethyl (racemate, 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.

2. The benzo(o)oxy(carbon / sulfur)azepine compound according to claim 1, characterized in that A compound selected from the following structures: 。 3. Use of the benzooxy(carbon / sulfur)azepine compound according to claim 1 in the preparation of monoamine oxidase B inhibitors.

4. The use of the benzooxy(carbon / sulfur)azepine compound according to claim 1 in the preparation of a pharmaceutical preparation, characterized in that: The pharmaceutical preparation is a pharmaceutical preparation for preventing and / or treating diseases related to monoamine oxidase B.

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

6. The use according to claim 5, characterized in that: The diseases related to monoamine oxidase B are one or more of Parkinson's disease, Alzheimer's disease, malignant tumors, depression, and anxiety.

7. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the benzooxy(carbon / sulfur)azepine compound or a pharmaceutically acceptable salt, cocrystal or solvate thereof.

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

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