CN119977904A - A benzooxy (carbon/sulfur) azaphene compound and its application in medicine - Google Patents

Abstract

The invention discloses a benzoxy (carbon/sulfur) aza-zepine compound and application thereof. The structural general formulas of the benzoxy (carbon/sulfur) azepine compounds are shown in the following formulas (I) and (II): Wherein A is selected from methyl (racemate, R configuration, S configuration), ethyl (racemate, R configuration, S configuration) or cyclopropyl, X ₁ is selected from oxygen, ammonia or sulfur, X ₂ is selected from methylene, oxygen or sulfur, R ₁ is selected from hydrogen, fluorine, chlorine, bromine or methyl, and R ₂ is selected from hydrogen, fluorine, chlorine, bromine or methyl. The benzoxy (carbon/sulfur) aza-zepine compound has good monoamine oxidase B inhibition activity and extremely high selectivity in monoamine oxidase family, and has good therapeutic effect in a mouse parkinsonism model.

Description

Benzoxy (carbon/sulfur) azepine compound and application thereof in medicine

Technical Field

The invention belongs to the field of medicines, and particularly relates to a benzo-oxy (carbon/sulfur) azepine compound and application thereof.

Background

Neurodegenerative diseases have long been considered one of the most mystery and challenging problems in the biomedical field. As the study of neurodegenerative diseases goes from descriptive phenomenological toward mechanistic analysis, it is becoming increasingly clear that their pathogenesis is often co-caused by multiple factors (genetic, environmental and endogenous). Common mechanisms include protein misfolding and aggregation, oxidative stress and free radical formation, dysregulated metal homeostasis, mitochondrial dysfunction and protein phosphorylation.

Parkinson's Disease (PD) is the second severe neurodegenerative disease following Alzheimer's Disease (AD). Studies have shown that monoamine oxidase B (MAO-B) is involved in the development and progression of parkinson's disease, and that inhibition of monoamine oxidase activity by drugs increases Dopamine (DA) levels, thereby improving symptoms in PD patients. Loss of DA is considered a pathological hallmark of parkinson's disease, and current therapeutic strategies are mainly focused on increasing the level of DA in the brain. Various drugs for alleviating symptoms of parkinson's disease have been developed and used clinically, wherein MAO-B inhibitors show better efficacy and more advantageous safety.

The existing MAO-B inhibitors used clinically can be divided into three generations, namely selegiline is an irreversible MAO-B inhibitor of generation 1. The chemical structure of the compound belongs to phenethylamine derivatives, and after in-vivo metabolism, metabolic products are amphetamine derivatives with sympathomimetic activity, and the metabolites with the sympathomimetic activity can increase the risks of heart diseases and hypertension of patients in the process of taking selegiline to treat PD. Because of the defects of poor selectivity, large adverse reaction and the like, the compound cannot be used as a first-choice medicament for treating PD. The generation 2 irreversible MAO-B selective inhibitor is rasagiline. The drug effect group is propargylamine, so that rasagiline can irreversibly inhibit monoamine oxidase to have stronger effect and better neuroprotection, and the metabolite is an inactive non-amphetamine substance, has small toxic and side effects, and is a common MAO-B inhibitor for clinically treating PD at present. However, rasagiline is still not an optimal drug for the treatment of PD due to its poor selectivity for MAO-B and irreversible inhibition of enzyme activity. The 3 rd generation MAO-B inhibitor is a reversible MAO-B selective inhibitor, namely, the safinamide. Unlike the traditional MAO-B inhibitor selegiline and rasagiline, the salfenamide has higher selectivity on MAO-B, reversible effect and safer clinical use. The safinamide also inhibits glutamate release and dopamine and serotonin reuptake. However, some patients may experience discomfort symptoms such as movement inconvenience, reduced sleep quality, or nausea after taking the drug. The types of selective MAO-B inhibitors in clinical application are very limited, and the diversity of treatment requirements is difficult to meet. Therefore, research and development of reversible, highly selective monoamine oxidase B inhibitors, i.e. third generation monoamine oxidase inhibitors, has become a hotspot in this field.

Disclosure of Invention

In view of the above-mentioned shortcomings of the prior art, the present invention provides a benzoxy (carbon/sulfur) azepine compound and its use in a medicament for preventing and/or treating a disease associated with monoamine oxidase B (Monoamine oxidase B). The benzoxy (carbon/sulfur) azepine compound has good monoamine oxidase B inhibition activity and shows ultra-high selectivity in monoamine oxidase family.

The structure general formula of the benzoxy (carbon/sulfur) azepine compound is shown as the following formulas (I) and (II):

Wherein:

A is selected from methyl (racemate, R configuration, S configuration), ethyl (racemate, R configuration, S configuration) or cyclopropyl, X ₁ is selected from oxygen, ammonia or sulfur, X ₂ is selected from methylene, oxygen or sulfur, R ₁ is selected from hydrogen, fluorine, chlorine, bromine or methyl, and R ₂ is selected from hydrogen, fluorine, chlorine, bromine or methyl.

Further, the benzoxy (carbon/sulfur) aza-zepine compound is selected from the following structures:

The invention relates to application of benzo-oxy (carbon/sulfur) azepine compounds in preparing pharmaceutical preparations.

The pharmaceutical preparation is used for preventing and/or treating monoamine oxidase B related diseases.

The monoamine oxidase B-related diseases include, but are not limited to, neurodegenerative diseases such as parkinson's disease, alzheimer's disease, malignant tumor, depression, anxiety, etc.

The invention also provides a pharmaceutical composition, the pharmaceutical composition comprises the benzoxy (carbon/sulfur) azepine compound or a pharmaceutically acceptable salt, co-crystal or solvate thereof.

The pharmaceutical composition further comprises pharmaceutically acceptable excipients and carriers.

The pharmaceutical composition is in the form of tablet, capsule, powder, granule, syrup, solution, oral liquid, spirit, aerosol, powder fog, injection, sterile powder for injection or suppository.

The pharmaceutical composition is administered via oral, intravenous, intramuscular or subcutaneous routes.

In particular, the benzoxy (carbon/sulfur) aza-compound can also exist in the medicament in the form of a solvate thereof, or a pharmaceutically acceptable salt thereof. The term "pharmaceutically acceptable salts" refers to salts of the compounds with acids or bases which are suitable for use as medicaments, including inorganic and organic salts. A preferred class of salts of the invention are those formed by benzooxy (carbon/sulfur) azepine and acids, and 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, phosphoric acid, and the like, 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 the like, and acidic amino acids such as aspartic acid, glutamic acid, and the like.

Specifically, the medicament further comprises a pharmaceutically acceptable carrier. A "pharmaceutically acceptable" ingredient is a substance that is suitable for use in humans and or animals without undue adverse side effects (such as toxicity, irritation, and allergic response), commensurate with a reasonable benefit/risk ratio. A "pharmaceutically acceptable carrier" is a pharmaceutically or food acceptable solvent, suspending agent or excipient for delivering a compound of the invention to an animal or human. The carrier may be a liquid or a solid. More specifically, pharmaceutically acceptable carriers are various pharmaceutically commonly used excipients and/or excipients, including, but not limited to, sugars (e.g., lactose, dextrose, and sucrose), starches (e.g., corn starch and potato starch), celluloses and derivatives thereof (e.g., sodium carboxymethyl cellulose, ethyl cellulose, and methyl cellulose), gum tragacanth powder, malt, gelatin, talc, solid lubricants (e.g., stearic acid and magnesium stearate), calcium sulfate, vegetable oils (e.g., peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and cocoa butter), polyols (e.g., propylene glycol, glycerol, sorbitol, mannitol, and polyethylene glycol), begonic acid, emulsifiers (e.g., tween/polyvinyl chloride castor oil), wetting agents (e.g., sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, isotonic saline solution, phosphate buffers, and the like, and the carriers may enhance the stability, activity, bioavailability, and the like of the formulation as desired.

Specifically, the medicine is tablet, capsule, powder, granule, syrup, solution, oral liquid, spirit, aerosol, powder fog, injection, sterile powder for injection or suppository.

The invention has the beneficial effects that the invention designs up>A series of molecular structures which are expected to have better inhibition activity and selectivity on MAO-B by utilizing up>A structure-based drug molecular design method according to the related report that the benzyloxy aryl derivatives can effectively inhibit the MAO-B and according to the slight difference between two targets of the MAO-A and the MAO-B, especially the difference between two receptor amino acid residues. Chemical synthesis and further biological activity tests show that among the benzoxy (carbon/sulfur) azepine compounds, when a seven-membered oxygen-containing ring exists, the benzoxy (carbon/sulfur) azepine compounds show very excellent activity on MAO-B, and show high selectivity inside the MAO family, and the series of compounds have the potential of treating diseases in which MAO-B participates in regulation. Further biological experiments confirm the superiority of some compounds in the treatment of parkinson's disease.

Drawings

FIG. 1 shows the reversibility of the compounds 17 and 19 according to the present invention.

FIG. 2 is a trace of an open field experiment.

Fig. 3 is the experimental results of 3 behavioral studies on MPTP-induced PD mice. The method comprises the following steps of (A) moving total distance of the mice, (B) entering frequency of central area of the mice, (C) residence time of central area of the mice, (D) moving average speed of the mice, (E) residence time of the mice on a rod, and (F) grasping experimental scores of the mice.

Note that all were administered by intraperitoneal injection. Statistical significance was analyzed using 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

The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

¹ H NMR spectra in all examples of the invention were determined using a 600M superconducting nuclear magnetic resonance spectrometer (AVANCE NEO 600), the chemical shifts being expressed in ppm using the tetramethylsilane internal standard (0.00 ppm). ¹ H NMR representation method: s=singlet, d=doublet, t=triplet, m=multiplet, br=broadened, m = multiple peak(s), br=the width of the strip to be widened. If coupling constants are provided, they are in Hz.

The thin layer chromatography silica gel plate uses a smoke table yellow sea HSGF254 or Qingdao GF254 silica gel plate, the specification of the silica gel plate used by the Thin Layer Chromatography (TLC) is 0.15-0.2 mm, and the specification of the thin layer chromatography separation and purification product is 0.4-0.5 mm.

Column chromatography uses yellow sea silica gel of 200-300 meshes as a carrier.

Example 1:

The synthetic route is as follows:

First step, synthesizing 3-fluorobenzyloxy-4-hydroxybenzoic acid methyl ester

Methyl 2, 4-dihydroxybenzoate (6.3 g, 1.2 equiv) was placed in a round bottom flask, 800-120 mL acetonitrile was used as solvent, sodium carbonate (11.95 g, 2.5 equiv) was added and stirred at room temperature for 15min, p-fluorobenzyl bromide (5 g, 1.2 equiv) was added and stirred for 2 min, then the temperature was raised to 78 ℃ and refluxed for 4h, and the reaction was monitored by thin layer chromatography TLC. After the reaction, the solvent was dried by spin-drying, and the target compound (4.25 g, yellow oil, yield 47.2%) was obtained by column chromatography.

Second step Synthesis of methyl 2- (2- ((tert-Butoxycarbonyl) amino) ethoxy) -4- ((3-fluorobenzyl) oxy) benzoate

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, 30-50 mL tetrahydrofuran was used as a solvent, stirred at 0 ℃ for 5 min, diisopropyl azodicarboxylate (1.77 g, 1.4 equiv), N ₂ was added dropwise, stirred at room temperature for 24h, and monitored by thin layer chromatography TLC. After the reaction, the solvent was dried by spin-drying, and the target compound (1.8 g, white oil, yield 68.7%) was obtained by extraction and column chromatography.

Third step, synthesis of methyl 2- (2-amino ethoxy) -4- (3-fluorobenzyl) oxy) benzoate

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

Fourth step, synthesis of 8- [ (3-fluorophenyl) methoxy ] -2,3,4, 5-tetrahydro-1, 4-benzoxazolin-5-one

Methyl 2- (2-aminoethoxy) -4- (3-fluorobenzyl) oxy) benzoate (1.1 g, 1 equiv) was placed in a round bottom flask and stirred at 0 ℃ with 10-20 mL methanol as solvent for 5min, sodium methoxide (1.4 g, 8 equiv) was added, stirred at room temperature for 9: 9 h, and monitored by thin layer chromatography TLC. After the reaction, the mixture was dried by spin-drying, and the target compound (0.7 g, white oil, yield: 71.4%) was obtained by extraction and column chromatography.

Fifth step, synthesis of 8- [ (3-fluorophenyl) methoxy ] -2,3,4, 5-tetrahydro-1, 4-benzoxazoline

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, aluminum chloride (0.49 g, 1.5 equiv) was added with 10-20mL tetrahydrofuran as solvent, stirred at-20℃for 5 min, lithium aluminum hydride (0.37 g, 4 equiv) was added, stirred for 2 min, and then warmed to 60℃with reflux for 6 h, and the reaction was monitored by thin layer chromatography TLC. After the reaction, it was quenched with 10% aqueous sodium hydroxide solution, the filtrate was dried by suction filtration, and the objective compound (0.5 g, white oil, yield 66.6%) was obtained by extraction and column chromatography.

Sixth step Synthesis of 2- {8- [ (3-fluorophenyl) methoxy ] -2,3,4, 5-tetrahydro-1, 4-benzoxazolin-4-yl } propanamide

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 stirred at room temperature for 15 min, 2-aminopropionamide (67.2 mg, 1.2 equiv), potassium iodide (6 mg, 0.1 equiv) were added and stirred for 2 min and then heated to 110℃and stirred under reflux for 4h, and the reaction was monitored by TLC. After the completion of the reaction, the objective compound (80 mg, white solid, yield 62.9%) was obtained by extraction and column chromatography.

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 = 6.8 Hz, 3H）.

Example 2:

Preparation procedure referring to example 1, methyl 2, 4-dihydroxybenzoate was converted to methyl 2, 5-dihydroxybenzoate, with the other conditions unchanged.

¹H NMR（600 MHz, Chloroform-d） δ（ppm）7.37-7.32（m, 1H）, 7.17（d, J = 7.5 Hz, 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）.

Example 3:

Preparation procedure referring to example 1, m-fluorobenzyl bromide was converted to p-fluorobenzyl bromide, with the other conditions unchanged.

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）.

Example 4:

preparation procedure referring to example 1, m-fluorobenzyl bromide was changed to p-fluorobenzyl bromide, methyl 2, 4-dihydroxybenzoate was changed to methyl 2, 5-dihydroxybenzoate, and the other conditions were unchanged.

¹H 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）.

Example 5:

The synthetic route is as follows:

First step Synthesis of 7-methoxy-2, 3, 4-5-tetrahydro-1H-benzazepan-1-one

6-Methoxy-3, 4-dihydronaphthalen-1 (2H) -one (1 g, 1 equiv) is placed in a round bottom flask, 10-20 mL of methanesulfonic acid is used as a solvent, 5-min is stirred at 0 ℃, sodium azide (1.1 g, 3 equiv) is slowly added, 18-H is stirred at room temperature, and the reaction condition is monitored by thin layer chromatography TLC. After the reaction, the solvent was dried by spin-drying, and the target compound (0.9 g, white solid, yield 83.3%) was obtained by extraction and column chromatography.

Second step, synthesis of 7-hydroxy-2, 3, 4-5-tetrahydro-1H-benzazepan-1-one

7-Hydroxy-2, 3, 4-5-tetrahydro-1H-benzazepan-1-one (0.9 g, 1 equiv) is placed in a three-neck flask in an N ₂ atmosphere, 10-20 mL of dichloromethane is used as a solvent, stirring is carried out at-78 ℃ for 5min, boron tribromide (4.71 g, 5 equiv) is dropwise added through a needle tube, the temperature is slowly increased to 0 ℃ for 4 hours of reaction, and the reaction condition is monitored by thin layer chromatography TLC. After the reaction, the mixture was quenched with saturated sodium carbonate aqueous solvent, and subjected to extraction and column chromatography to give the objective compound (0.75 g, white solid, yield 90.3%)

Third step Synthesis of 7- ((4-fluorobenzyl) oxy) -2,3, 4-5-tetrahydro-1H-benzazepan-1-one

Referring to example 1, m-fluorobenzyl bromide was converted to p-fluorobenzyl bromide, methyl 3-fluorobenzyloxy-4-hydroxybenzoate was converted to 7-hydroxy-2, 3, 4-5-tetrahydro-1H-benzazepan-1-one, with the other conditions unchanged.

Synthesis of 2- (7- ((4-fluorobenzyl) oxy) -1,3, 4-5-tetrahydro-2H-benzazepan-2-yl) propanamide

Referring to example 1, 8- [ (3-fluorophenyl) methoxy ] -2,3,4, 5-tetrahydro-1, 4-benzoxazolin-5-one was exchanged for 7- ((4-fluorobenzyl) oxy) -2,3, 4-5-tetrahydro-1H-benzazepan-1-one, with the other conditions unchanged.

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, 5.3 Hz, 1H）, 1.11（d, J = 6.9 Hz, 3H）.

Example 6:

Preparation procedure referring to example 5, p-fluorobenzyl bromide was exchanged for m-fluorobenzyl bromide, with the other conditions unchanged.

¹H 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）.

Example 7:

Preparation procedure referring to example 1, the m-fluorobenzyl bromide was exchanged for m-methylbenzyl bromide, with the other conditions unchanged.

¹H NMR（600 MHz, Chloroform-d） δ（ppm）7.25-7.17（m, 2H）, 7.14（d, J = 7.5 Hz, 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）.

Example 8:

Preparation procedure referring to example 5, p-fluorobenzyl bromide was exchanged for m-methylbenzyl bromide, with the other conditions unchanged.

¹H 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）.

Example 9:

preparation procedure referring to example 1, m-fluorobenzyl bromide was converted to p-methylbenzyl bromide with the other conditions unchanged.

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）.

Example 10:

Preparation procedure referring to example 1, the m-fluorobenzyl bromide was exchanged for benzyl bromide, with the other conditions unchanged.

¹H 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）.

Example 11:

preparation procedure referring to example 5, p-fluorobenzyl bromide was exchanged for benzyl bromide, with the other conditions unchanged.

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）.

Example 12:

preparation procedure referring to example 5, p-fluorobenzyl bromide was exchanged for 1-bromo-4- (bromomethyl) -2-fluorobenzene, with the other conditions unchanged.

¹H 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 = 22.7 Hz, 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）.

Example 13:

Preparation procedure referring to example 1, m-fluorobenzyl bromide was exchanged for 1-bromo-4- (bromomethyl) -2-fluorobenzene, with the other conditions unchanged.

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）.

Example 14:

Preparation procedure referring to example 1, the conversion of m-fluorobenzyl bromide to 4- (bromomethyl) -1, 2-dichlorobenzene was carried out under otherwise unchanged conditions.

¹H 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 = 22.7 Hz, 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）.

Example 15:

preparation procedure referring to example 1, the conversion of m-fluorobenzyl bromide to 4- (bromomethyl) -1, 2-difluorobenzene was carried out under otherwise unchanged conditions.

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 = 5.0 Hz, 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）.

Examples 16 to 19:

chiral resolution was performed on example 3 and example 15, respectively, to obtain the products of examples 16 to 19.

The profile parameters for the products of examples 16-19 are shown in the following table:

Example 20:

Preparation procedure referring to example 5, p-fluorobenzyl bromide was exchanged for 4- (bromomethyl) -1, 2-difluorobenzene, with the other conditions unchanged.

¹H 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）.

Example 21:

preparation procedure referring to example 1, the conversion of m-fluorobenzyl bromide to m-chlorobenzyl bromide was unchanged with other conditions.

¹H 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）.

The inhibitory activity of the compounds obtained in the above examples on MAO-A and MAO-B was measured as follows:

MAO-A/B enzyme was diluted 800-fold with sodium phosphate buffer (0.05M, pH=7.4) for use. The weighed (about 5 mg) test compound was dissolved in DMSO solution and diluted to the desired concentration with sodium phosphate buffer (DMSO content in the prepared solution was controlled to be no more than 1%). 80 mu L of diluted MAO-A or MAO-B enzyme and 20 mu L of compounds with different concentration gradients are sequentially added into up>A black 96-well plate, wherein 20 mu L of sodium phosphate buffer solution is added into up>A blank group, and then 15 mins is incubated in up>A 37 ℃ incubator, so that the compound to be tested is combined with the MAO-B enzyme, and the combination of the MAO-A/B enzyme and substrate tyramine is effectively inhibited. 15 After the mins incubation period, 100. Mu.L of substrate mixed solution (1100. Mu.L of sodium phosphate buffer, 500. Mu.L of tyramine, 400. Mu.L of horseradish peroxidase, 20. Mu.L of complex Red reagent) was added to each well for reaction, and the total volume of the whole reaction system was 200. Mu.L. After the completion of the substrate addition, the amount of H ₂O₂ generated by the reaction of MAO-B with tyramine was calculated by measuring the fluorescence intensity (30 min by scanning at excitation light wavelength 545 nm and absorption light wavelength 590 nm at a time interval of 2 min) in an enzyme-labeled instrument (FLX 800, bio-Tek Instruments, inc., synergy, HI, USA) immediately and fitting the data read for each time period by using GRAPHPAD PRISM 9.5.5 software, thereby calculating the IC ₅₀ value of the compound.

The results of the biological activity experiments of compounds 1-21 and positive controls on MAOs are shown in the following table.

Long-term use of irreversible MAO-B inhibitors induces up-regulation of diamine oxidase compensatory gene expression, resulting in ineffective decrease of GABA levels in astrocytes, thus limiting the application potential of irreversible MAO-B inhibitors in neuroprotection. Based on this, rasagiline, an irreversible inhibitor, was selected as a positive control and the mode of action of compound 17, compound 19 and hMAO-B was assessed.

The test procedure is essentially identical to the test hMAO-A and/or hMAO-B IC ₅₀ procedure, by first incubating with a high concentration of inhibitor (50 XIC ₅₀) for a period of time, allowing the enzyme to bind the compound sufficiently, almost completely, and then diluting rapidly one hundred times with buffer. For reversible inhibitors, the enzyme activity slowly recovers with the extension of the time after dilution, while for irreversible inhibitors, the enzyme activity does not recover.

Fig. 1 shows the reversibility of compound 17 and compound 19. From FIG. 1 it can be seen that the test results are consistent with the reported results in that rasagiline acts in an irreversible manner with hMAO-B. As the slope of the curves of compound 17 and compound 19 increases toward the control group with the extension of the test time, the inhibition rate decreases, and the enzyme activity slowly recovers with the extension of the time after dilution, indicating that the modes of action of compound 17, compound 19 and hMAO-B are reversible combinations.

Since PD patients often have motor dysfunction such as bradykinesia, muscle stiffness, resting tremor, and gait disturbance, the quality of life of the patient is severely reduced. Therefore, assessing whether a compound improves motor dysfunction is an important indicator of pharmacodynamic evaluation. 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (1-Methyl-4-phenyl-1, 2,3,6-tetrahydropyridine, MPTP) is a neurotoxin with good lipid solubility which can rapidly cross the blood brain barrier into the nervous system. In glial cells, MPTP is metabolized by MAO-B to form MPDP+ intermediates, which are further oxidized to form the toxic metabolite MPP+, resulting in damage and death of dopaminergic neurons in SNpc, inducing typical motor symptoms of PD. Based on the characteristics of compound 17 and compound 19, such as excellent MAO-B inhibitory activity, pharmacodynamics thereof was studied intensively in the MPTP-induced PD subacute mouse model.

C57BL/6J mice (mice age: 2 months, body weight: 25.+ -.2 g) purchased from Henan Seebeck Biotechnology Co., ltd.) were randomly divided into 7 groups (n=6/group) consisting of a blank group, an MPTP group, an MPTP+sand fenamide (30 mg/kg) group, an MPTP+ compound 17 (10 mg/kg) group, an MPTP+ compound 17 (30 mg/kg) group, an MPTP+ compound 19 (10 mg/kg) group, and an MPTP+ compound 19 (30 mg/kg) group. The groups were administered by intraperitoneal injection, wherein the blank and MPTP groups were given equal volumes of vehicle by intraperitoneal injection. 30 After min, MPTP, compound and safinamide groups were intraperitoneally injected with MPTP solution (20 mg/kg), and the blank group was intraperitoneally injected with an equal volume of physiological saline. The above operation was repeated for seven days. The behavioral performance of the mice was evaluated on day eight by open field experiments, stick rotation experiments, grip experiments. The specific behavioural experimental method is as follows:

(1) The open field experiment is mainly used for evaluating the autonomous movement condition of the mice. The experiment was performed in dark conditions and independent tests were performed after mice were acclimatized to dark conditions for half an hour. Mice were placed in an open field experimental set-up (0.4x0.4x0.5 m ³) and were free to move 5 min. The area is divided by software, and the data such as the movement track and the movement time of the mice are collected by a camera for analysis.

(2) The rod rotation test aims at detecting the movement ability of the limbs of the mice. Mice adapt to instruments and environments one day in advance. In the experiment, the mice are placed on a horizontal rotating rod, the instrument is started in a uniform acceleration mode, the rotating speed is increased from 5 rpm to 40 rpm, and the instrument is stopped after 3 minutes. From the time the mice were able to rest smoothly on the rotating rod until they were dropped from the rotating rod or left for more than 3 minutes. The residence time of each mouse on the rotating rod was recorded and 3 experiments were repeated for each mouse.

(3) The grip test mainly evaluates the muscle relaxation performance and motor coordination of mice. The tail of the mouse was lifted so that its front paw grasped the wire, which was 30 cm long and 30 cm away from the ground, and then the tail was released to observe the mouse's grip. The grip was scored according to the following criteria:

The device comprises a front claw, a tail, a device and a tail, wherein the front claw is hung on a metal wire, the front claw is used for climbing upwards after grasping the metal wire, the front claw can be used for grasping the metal wire by one or two rear claws after grasping the metal wire, the tail is wound on the metal wire, and the tail reaches the tail end of the metal wire and escapes from the device.

FIG. 2 is a trace diagram of open field experiment, FIG. 3 is the experimental results of 3 behavioral studies on MPTP-induced PD mice (A) total distance of movement of mice, (B) number of times of entrance of central region of mice, (C) residence time of central region of mice, (D) average velocity of movement of mice, (E) residence time on rod, and (F) score of holding experiment of mice. Compared with the blank group, the on-stick time and the grip score and the open field experimental value of the MPTP group mice are obviously reduced, which indicates that the PD mice have obvious defects in muscle functions and exercise capacity. After treatment with the salfenamide (30 mg/kg), the various behavioral data parameters of mice are significantly improved, however, compound 17 or compound 19 can significantly improve the various behavioral indexes of PD model mice at an administration dose of 10 mg/kg, and the improvement effect is close to the level of the blank group. In conclusion, compound 17, compound 19, at low doses, have unique advantages in improving motor dysfunction in MPTP-induced PD mouse models, especially in improving muscle relaxation and improving motor coordination.

In summary, the invention designs up>A series of molecular structures with better inhibition activity and selectivity to MAO-B by using up>A structure-based drug molecular design method according to the slight difference between two targets of MAO-A and MAO-B, especially the difference between two receptor amino acid residues. Chemical synthesis and further biological activity tests showed that among the benzoxy (carbon/sulfur) azepine compounds, when a seven membered oxygen containing ring is present, they show very excellent activity towards MAO-B and high selectivity inside the MAO family. In the examples listed herein, more than half of the compounds had better inhibitory activity against MAO-B as well as selectivity than the positive control drug, safinamide. In addition, the invention also carries out a thin jump experiment on two representative compounds, and the experimental results show that the series of compounds and hMAO-B act in a reversible combination mode. Compound 17, compound 19 were able to significantly improve MPTP-induced motor dysfunction and muscle function impairment in mice at low doses in the mouse behavioural study. The findings further prove that the series of compounds have great potential in treating diseases involving MAO-B regulation, provide new ideas and effective drug candidate molecules for treating related diseases, and are expected to play an important role in future clinical application.

Although embodiments of the present 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 therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims (8)

1. A benzooxy (carbon/sulfur) azaphene compound, characterized in that its general structural formula is as shown in the following formulas (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 it is selected from the following compounds: . 3. Use of the benzooxy(carbon/sulfur)azepine compound according to claim 1 in the preparation of monoamine oxidase B inhibitors. 4. 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.