Compound with anti-neuroinflammation activity and selective butyrylcholine esterase inhibition activity

Abstract

The invention discloses a compound which has anti-neuroinflammation activity and can selectively inhibit butyrylcholine esterase activity, and belongs to the technical field of medicinal chemistry. The compound and the pharmaceutical composition containing the compound have good anti-neuroinflammatory activity and selective butyrylcholine esterase inhibition activity, and have important application prospects in treatment of neurodegenerative diseases such as Alzheimer's disease.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology and relates to a novel class of compounds with dual activities. These compounds can efficiently inhibit butyrylcholinesterase activity and significantly suppress neuroinflammatory responses, and can be used for the prevention and / or treatment of neurodegenerative diseases, especially Alzheimer's disease. Background Technology

[0002] Alzheimer's disease (AD) is a common neurodegenerative disease affecting older adults, characterized by progressive memory loss and cognitive impairment. This disease poses a serious threat to the health and well-being of the elderly and places a heavy burden on healthcare systems. Therefore, conquering AD is considered a major global medical challenge.

[0003] The pathogenesis of Alzheimer's disease (AD) is extremely complex, and the scientific community currently has several hypotheses, including cholinergic damage, β-amyloid (Aβ) cascade, abnormal phosphorylation of Tau protein, and neuroinflammation. However, no single theory can fully explain the entirety of the disease. Most clinically used anti-AD drugs target a single mechanism. For example, donepezil, galantamine, and rivastigmine are specific cholinesterase inhibitors that improve neurotransmission by increasing the concentration of acetylcholine in the brain; memantine is an NMDA receptor antagonist that reduces excitotoxicity by regulating glutamate activity. In recent years, monoclonal antibody drugs targeting Aβ, such as lecanemab and donanemab, have been approved for marketing, slowing disease progression by clearing amyloid plaques in the brain. However, except for memantine, which can be used for moderate to severe patients, most drugs are only suitable for patients with mild dementia. These drugs cannot completely cure the disease and generally have significant side effects, making long-term use unsuitable. In particular, some antibody drugs may cause serious risks such as cerebral edema or cerebral hemorrhage.

[0004] Throughout the long course of Alzheimer's disease (AD), many patients with severe AD still lack effective treatment options. Studies have found that in the early stages of AD, acetylcholinesterase (AChE) primarily hydrolyzes acetylcholine (ACh). However, in the mid-to-late stages of AD, AChE levels in the brain decrease to 55%–67% of normal, and its enzyme activity is essentially lost. Simultaneously, butyrylcholinesterase (BuChE) activity increases to approximately 1.2 times its original level. In vivo experiments have demonstrated that selective BuChE inhibitors can improve cognitive function in animals without the peripheral nervous system side effects associated with AChE inhibitors. Therefore, for the treatment of moderate to severe AD, selectively inhibiting BuChE in the brain, rather than AChE, has become a key therapeutic strategy.

[0005] Furthermore, the "neuroinflammatory hypothesis" has received increasing support from research in recent years. A study published in *Nature Medicine* found that some elderly individuals, despite having amyloid plaques in their brains, did not develop Alzheimer's disease. The interaction between microglia-mediated neuroinflammation and amyloid protein is considered a key factor driving the spread of Aβ pathology to tau protein, ultimately leading to brain damage and cognitive impairment. Intervening in the neuroinflammatory process holds promise for reversing or slowing the accumulation of pathological tau protein in the brain, thereby preventing dementia and benefiting early-stage AD patients. Therefore, inhibiting neuroinflammation may become a new treatment option for mild Alzheimer's disease.

[0006] In conclusion, compared with single-mechanism drugs, the combined treatment strategy of selectively inhibiting BuChE and anti-neuroinflammatory drugs is more reasonable and has greater development value.

[0007] Current Alzheimer's disease (AD) treatments still face limitations in terms of choice. Long-term medication often leads to cumulative side effects or drug resistance, making subsequent treatment options more difficult. Furthermore, there are relatively few selective BuChE inhibitors available, and many, while inhibiting BuChE, also have some effect on AChE, lacking high selectivity. Therefore, developing novel, highly effective selective butyrylcholinesterase inhibitors with anti-neuroinflammatory activity is of significant scientific and clinical value for improving the current state of Alzheimer's disease treatment. Summary of the Invention

[0008] To address the shortcomings of existing technologies, this invention first provides a class of compounds that possess both anti-neuroinflammatory activity and the ability to selectively inhibit butyrylcholinesterase activity. Secondly, it provides screening results of these compounds' inhibitory activity on butyrylcholinesterase and their inhibitory activity on microglia inflammation, as well as their application in the preparation of drugs for the prevention and / or treatment of neurodegenerative diseases such as Alzheimer's disease.

[0009] To achieve the above-mentioned objectives, this invention has screened and obtained a class of compounds having the structure shown in formula (I) below, and pharmaceutically acceptable salts thereof:

[0010] in: R1 is hydrogen or a C1-C4 alkyl group; R2 is a substituted amino group or a 5- or 6-membered nitrogen heterocyclic group.

[0011] More specifically, R1 is hydrogen, methyl, ethyl, propyl, n-butyl, or tert-butyl; R2 is N,N-dimethylamino, N-ethyl-N-methylamino, N,N-diethylamino, N,N-diisopropylamino, N,N-di(2-chloroethyl)amino, pyrrolidone-1-yl, piperidin-1-yl, or morpholino-4-yl.

[0012] That is, the compound with the structure shown in formula (I) of this invention can be one of the following compounds: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

[0013] Preferably, R1 can be hydrogen, methyl, ethyl, propyl, n-butyl or tert-butyl; R2 can be N,N-dimethylamino, N-ethyl-N-methylamino, N,N-diisopropylamino, N,N-di(2-chloroethyl)amino, pyrrolidone-1-yl or piperidin-1-yl.

[0014] More preferably, R1 can be hydrogen, methyl, ethyl, n-butyl or tert-butyl; R2 can be N,N-dimethylamino, N-ethyl-N-methylamino, N,N-diisopropylamino, pyrrolidone-1-yl or piperidin-1-yl.

[0015] More preferably, R1 can be hydrogen, methyl, n-butyl or tert-butyl; R2 can be N,N-diisopropylamino, pyrrolidone-1-yl or piperidin-1-yl.

[0016] In particularly preferred compounds, R1 is methyl or n-butyl, and R2 is piperidin-1-yl.

[0017] The "pharmaceutically acceptable salt" as described in this invention refers to a salt that, within the scope of reasonable medical judgment, is suitable for contact with human and animal tissues without undue toxicity, irritation, allergic reactions, or other problems or complications.

[0018] Therefore, compounds with the structure shown in formula (I) may include all pharmaceutically acceptable salts thereof, including but not limited to acid addition salts formed with inorganic or organic acids, and base addition salts formed with inorganic or organic bases.

[0019] Among them, acid addition salts include, but are not limited to: salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, and phosphoric acid; or salts formed with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and camphorsulfonic acid; and salts formed with amino acids (such as glutamic acid and aspartic acid).

[0020] Among them, the base addition salts include, but are not limited to: ammonium salts; alkali metal salts (such as sodium salts, potassium salts, lithium salts); alkaline earth metal salts (such as calcium salts, magnesium salts); salts formed with organic bases (such as primary amines, secondary amines, tertiary amines); and quaternary ammonium salts generated by the reaction of the basic nitrogen-containing group in the compound of formula (I) with an alkylating agent (such as lower alkyl halides, aralkyl halides), wherein the alkylating agent includes, but is not limited to, dialkyl sulfate esters (such as dimethyl sulfate, diethyl sulfate), long-chain alkyl halides (such as decyl chloride, lauryl chloride, stearyl chloride), etc.

[0021] The compound with the structure shown in formula (I) of this invention can be prepared by various feasible methods. For example, it can be prepared by following the synthetic route shown in the following equation: benzoic acid with different R1 substitutions at position 4 reacts with sulfoxide under N,N-dimethylformamide catalysis to obtain a first intermediate; o-hydroxyanisole reacts with formyl chloride with different R2 substitutions in the presence of catalyst 4-dimethylaminopyridine and acid-binding agent N,N-diisopropylethylamine to obtain a second intermediate; the first intermediate and the second intermediate undergo Friedel-Crafts acylation under AlCl3 catalysis to obtain a third intermediate; finally, the third intermediate undergoes demethylation under boron tribromide to obtain the compound with the structure shown in formula (I).

[0022]

[0023] in: R1 is hydrogen or a C1-C4 alkyl group; R2 is a substituted amino group or a 5- or 6-membered nitrogen heterocyclic group.

[0024] It should be noted that the above preparation method is only a preferred preparation method for the compound described in this invention, and is not intended to limit the preparation method of the compound described in this invention.

[0025] Using donepezil as a positive control, this invention tested the inhibitory activity of the synthesized compounds against AChE and BuChE. The results showed that most compounds exhibited strong BuChE inhibitory activity and low AChE inhibitory activity, demonstrating high selectivity for BuChE. In particular, compound JOPQ showed nanomolar-level high BuChE inhibitory activity.

[0026] This invention also demonstrates that the compound can inhibit LPS-induced microglial inflammatory responses, exhibiting good anti-neuroinflammatory activity.

[0027] The corresponding behavioral studies and biochemical index detection of this invention further corroborate that it is a highly selective butyrylcholinesterase inhibitor with anti-neuroinflammatory activity.

[0028] Therefore, the present invention also provides the use of the compound in the preparation of a medicament for the prevention and / or treatment of neurodegenerative diseases. The neurodegenerative diseases may include, but are not limited to, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), or spinocerebellar ataxia, preferably Alzheimer's disease or Parkinson's disease.

[0029] In addition, the present invention also provides a pharmaceutical composition comprising the compound described herein or a pharmaceutically acceptable salt thereof as an active ingredient, and one or more pharmaceutically acceptable carriers or excipients.

[0030] Compared with existing technologies, the compounds of this invention possess a variety of favorable in vitro activities and exhibit multiple similarities to the pathogenesis of neurodegenerative diseases such as Alzheimer's disease, enabling treatment of Alzheimer's disease through various pathways. In particular, the compounds of this invention can treat mild, moderate, and severe Alzheimer's disease by inhibiting neuroinflammation and selectively inhibiting butyrylcholinesterase activity. Therefore, the compounds of this invention show great promise for application in the preparation of anti-Alzheimer's disease drugs. Attached Figure Description

[0031] Figure 1 This is a Lineweaver-Burk double reciprocal plot of the interaction between compound JOPQ and BuChE.

[0032] Figure 2 This describes the effect of compound JOPQ on the trajectory of mice in a novel object recognition experiment.

[0033] Figure 3 The effect of compound JOPQ on the trajectory of mice in the water maze experiment. Implementation

[0034] The embodiments of the present invention will now be described in detail with reference to the accompanying drawings and specific examples. It should be noted that the embodiments are merely illustrative and are intended to provide a thorough understanding of the technical solutions of the present invention and to provide guidance for those skilled in the art to implement and apply the present invention. It should be understood that these descriptions do not constitute any limitation on the scope of protection of the present invention.

[0035] Unless otherwise expressly stated, the production processes, experiments, tests or analysis methods involved in the embodiments of the present invention are all considered to be conventional methods known to those skilled in the art, and only need to be implemented in accordance with conventional conditions or relevant product instructions. The steps and names involved are also generally clear and unambiguous in the art.

[0036] The instruments, equipment, raw materials, reagents, or samples used in the embodiments are not subject to any special restrictions on their source. They are all conventional products that can be purchased through regular commercial channels or prepared by known methods, and their source does not have a substantial impact on the implementation results of the present invention.

[0037] Unless otherwise expressly defined, the scientific and technical terms used in this invention have the meanings commonly understood by one of ordinary skill in the art. In case of any conflict, the definitions in this specification shall prevail.

[0038] The terms “comprising,” “including,” “having,” etc., used in this invention should be understood as open-ended, meaning “including but not limited to.” The term “and / or” includes any and all combinations of one or more of the associated listed items. Quantitative terms such as “a,” “one,” etc., do not exclude multiples; “multiple” or “a variety” refers to quantities greater than or equal to two.

[0039] The terms "preferred", "better", and "exemplary" used in this invention are only used to describe specific solutions or effects and are not intended to limit the necessary scope of the solution or the scope of protection.

[0040] This invention relates to the description of numerical parameters (such as quantity, concentration, temperature, time, etc.), and it should be understood that reasonable deviations naturally exist due to measuring instruments, operational errors, statistical fluctuations, etc. The range of such deviations should be within limits acceptable to those skilled in the art based on common sense.

[0041] This invention relates to the description of numerical parameters (such as quantity, concentration, temperature, time, etc.), and it should be understood that reasonable deviations naturally exist due to measuring instruments, operational errors, statistical fluctuations, etc. The range of such deviations should be within limits acceptable to those skilled in the art based on common sense.

[0042] The following embodiments of the present invention specifically provide a class of compounds that possess both anti-neuroinflammatory activity and selective inhibition of butyrylcholinesterase activity, which are compounds having the structure shown in formula (I) below, and pharmaceutically acceptable salts thereof:

[0043] in: R1 is hydrogen or a C1-C4 alkyl group; R2 is a substituted amino group or a 5- or 6-membered nitrogen heterocyclic group.

[0044] The term "pharmaceutically acceptable salt" specifically refers to a conventional acid addition salt that can retain the biological activity and properties of the structure of formula (I) and form with a suitable nontoxic inorganic or organic acid, and these salts are also included within the scope of this invention.

[0045] More specifically, the compound may be a compound corresponding to the structural formula indicated by the number in Table 1.

[0046]

[0047]

[0048] The compounds with the structure shown in formula (I) of this invention can be prepared by the following exemplary synthetic route:

[0049] S1: Benzoic acid with different R1 substitutions at the 4-position is mixed with excess thionyl chloride and subjected to a substitution reaction under reflux for 5–10 h in the presence of a catalytic amount of N,N-dimethylformamide. The excess thionyl chloride is removed by vacuum distillation to obtain the corresponding 4-substituted benzoyl chloride as the first intermediate for the next step of the reaction.

[0050] S2: Dissolve o-hydroxyanisole in anhydrous dichloromethane, add acid-binding agent N,N-diisopropylethylamine and catalyst 4-dimethylaminopyridine, slowly add R2-substituted formyl chloride, stir overnight at room temperature to prepare the esterified product as the second intermediate.

[0051] S3: Dissolve the first and second intermediates in anhydrous dichloromethane, add Lewis acid catalyst AlCl3, stir and mix under ice bath, transfer to room temperature and continue stirring to carry out Friedel-Crafts acylation reaction, pour the reactants into ice water to quench the reaction, and extract to obtain the third intermediate.

[0052] S4: The third intermediate was dissolved in anhydrous dichloromethane, boron tribromide was added dropwise under ice bath, and the mixture was stirred overnight at room temperature to carry out the demethylation reaction. After the reaction was terminated with ice water, the final target product was obtained by extraction with ethyl acetate.

[0053] It should be noted that the above description is merely an exemplary scheme. The reaction solvent, temperature, time, catalyst, and post-treatment method can all be appropriately adjusted based on common knowledge in the field, and all such adjustments fall within the protection scope of this invention.

[0054] The compounds involved in the embodiments of the present invention have significant inhibitory effects and high selectivity on butyrylcholinesterase, and have good anti-neuroinflammatory activity. Therefore, they can be used in the preparation of drugs for the prevention or treatment of neurodegenerative diseases such as Alzheimer's disease. Example

[0055] Example 1

[0056] Weigh 6g of benzoic acid (37.45mmol) and add it to a reaction flask. Then add 15mL of thionyl chloride and 0.5mL of N,N-dimethylformamide in sequence. Reflux for 8h. Remove thionyl chloride by rotary evaporation to obtain crude benzoyl chloride.

[0057] 4 mL of o-hydroxyanisole (36.37 mmol), 0.44 g of 4-dimethylaminopyridine (3.637 mmol), and 9.5 mL of N,N-diisopropylethylamine (54.55 mmol) were sequentially added to a flask, dissolved in 20 mL of anhydrous dichloromethane, and stirred thoroughly for 10 min. Then, 5 mL of N,N-dimethylcarbamoyl chloride (54.55 mmol) was added, and the mixture was reacted at room temperature for 18 h. The reaction progress was monitored by TLC. After the reaction was completed, the solvent was removed by rotary evaporation, and the mixture was separated by silica gel column chromatography to obtain 2-methoxyphenyl N,N-dimethylcarbamate.

[0058] 0.89 mL of benzoyl chloride (7.68 mmol), 1 g of N,N-dimethylcarbamate 2-methoxyphenyl ester (5.1 mmol), 20 mL of anhydrous dichloromethane, and 1.54 g of anhydrous aluminum trichloride (11.52 mmol) were added sequentially to a flask. The mixture was stirred at 0–4 °C for 10 min, then allowed to react at room temperature. The reaction was monitored by TLC. The reaction was terminated by adding the reaction solution dropwise to icy dilute hydrochloric acid. The mixture was extracted three times with dichloromethane, and the organic phases were combined. The mixture was dried over anhydrous sodium sulfate overnight, the solvent was removed by rotary evaporation, and the product was separated by silica gel column chromatography to obtain 3-(N,N-dimethylcarbamoyloxy)-4-methoxybenzophenone.

[0059] 1 g of 3-(N,N-dimethylcarbamoyloxy)-4-methoxybenzophenone (3.34 mmol) was weighed and dissolved in 10 mL of anhydrous dichloromethane. 1.28 mL of boron tribromide (13.36 mmol) was added at 0–4 °C. After 10 min, the mixture was transferred to room temperature, and the reaction was monitored by TLC. The reaction was terminated by adding 10 mL of ice water. The mixture was extracted three times with ethyl acetate, and the organic phases were combined. The mixture was dried over anhydrous sodium sulfate overnight, the solvent was removed by rotary evaporation, and the product was separated by silica gel column chromatography to give 3-(N,N-dimethylcarbamoyloxy)-4-hydroxybenzophenone (1OJQ) as a white solid, with a yield of 62%.

[0060] mp: 151.5-152.7℃; ESI-MS: m / z 284.81 (M-1), C 16 H 15 O4N (285.10).

[0061] 1 H NMR (600 MHz, DMSO-d6) δ 10.67 (s, 1H), 7.69 – 7.65 (m, 2H), 7.65– 7.61 (m, 1H), 7.54 (s, 1H), 7.52 (dd, J = 8.5, 2.2 Hz, 1H), 7.42 (d, J=2.2 Hz, 1H), 7.04 (d, J = 8.4 Hz, 1H), 6.54 (s, 1H), 3.04 (s, 3H), 2.89 (s, 3H).

[0062] Example 2

[0063] Except for replacing N,N-dimethylcarbamoyl chloride with N,N-diethylcarbamoyl chloride, the same procedure as in Example 1 was followed to prepare 3-(N,N-diethylcarbamoyloxy)-4-hydroxybenzophenone (1OYQ) as a white solid with a yield of 67%.

[0064] mp: 115.1-118.5℃; ESI-MS: m / z 314.17 (M+1) + C 18 H 19 NO4 (313.4).

[0065] 1 H NMR (600 MHz, DMSO-d6) δ 10.70 (s, 1H), 7.69 – 7.66 (m, 2H), 7.63(d, J = 7.4 Hz, 1H), 7.55 (t, J = 7.6 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.42 (d, J = 2.2 Hz, 1H), 7.03 (d, J = 8.5 Hz, 1H), 3.43 – 3.39 (m, 2H), 3.28 (q, J = 7.2 Hz, 2H), 1.15 (dt, J = 63.0, 7.2 Hz, 6H).

[0066] Example 3

[0067] Except for replacing N,N-dimethylcarbamoyl chloride with N-ethyl-N-methylcarbamoyl chloride, the same procedure as in Example 1 was followed to prepare 3-(N-ethyl-N-methylcarbamoyloxy)-4-hydroxybenzophenone (1ONQ) as a white solid with a yield of 59%.

[0068] mp: 133.6-134.2℃; ESI-MS: m / z 300.20 (M+1) + C 17 H 17O4N (299.12).

[0069] 1 H NMR (600 MHz, DMSO-d6) δ 10.70 (s, 1H), 7.65 (d, J = 21.2 Hz, 3H), 7.53 (d, J = 24.2 Hz, 3H), 7.42 (s, 1H), 7.12 – 6.96 (m, 1H), 3.29 (s, 2H), 2.95 (d, J = 82.0 Hz, H), 1.14 (d, J = 59.9 Hz, 3H).

[0070] Example 4

[0071] Except for replacing N,N-dimethylcarbamoyl chloride with N,N-diisopropylcarbamoyl chloride, the same procedure was followed as in Example 1 to prepare 3-(N,N-diisopropylcarbamoyloxy)-4-hydroxybenzophenone (1OIQ) as a white solid with a yield of 55%.

[0072] mp: 148.1-150.1℃; ESI-MS: m / z 342.26 (M+1) + C 20 H 23 NO4 (341.4).

[0073] 1 H NMR (600 MHz, DMSO-d6) δ 10.68 (s, 1H), 7.68 (d, J = 7.0 Hz, 2H), 7.64 (t, J = 7.4 Hz, 1H), 7.54 (t, J = 7.6 Hz, 2H), 7.49 (dd, J = 8.4, 2.2Hz, 1H), 7.42 (d, J = 2.1 Hz, 1H), 7.02 (d, J = 8.4 Hz, 1H), 4.04 – 3.93 (m,2H), 1.24 (d, J = 35.2 Hz, 12H).

[0074] Example 5

[0075] Except for replacing N,N-dimethylcarbamoyl chloride with N,N-di(2-chloroethyl)carbamoyl chloride, the same procedure was followed as in Example 1 to prepare 3-[N,N-di(2-chloroethyl)carbamoyloxy]-4-hydroxybenzophenone (100Q) as a white solid with a yield of 68%.

[0076] mp: 154.6-155.9℃; ESI-MS: m / z 382.11 (M), C 18 H 17 C l2 NO4 (382.2).

[0077] 1 H NMR (600 MHz, DMSO-d6) δ 10.84 (s, 1H), 7.67 (d, J = 7.5 Hz, 2H), 7.64 (t, J = 7.4 Hz, 1H), 7.54 (s, 1H), 7.51 (s, 1H), 7.05 (d, J = 8.4 Hz,1H), 6.53 (s, 2H), 3.92 (t, J = 6.6 Hz, 2H), 3.78 (dt, J = 10.5, 6.5 Hz, 4H), 3.67 (t, J = 6.5 Hz, 2H).

[0078] Example 6

[0079] Except for replacing N,N-dimethylcarbamoyl chloride with 1-piperidinylcarbamoyl chloride, the same procedure as in Example 1 was followed to prepare 3-(1-piperidinylcarbamoyloxy)-4-hydroxybenzophenone (1OPQ) as a white solid with a yield of 72%.

[0080] mp: 171.2-172.9℃; ESI-MS: m / z 325.26 (M), C 19 H 19 O4N (325.13).

[0081] 1 H NMR (600 MHz, DMSO-d6) δ 10.69 (s, 1H), 7.68 (d, J = 1.3 Hz, 1H),7.65 – 7.62 (m, 1H), 7.54 (s, 1H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.42 (d,J = 2.2 Hz, 1H), 7.03 (d, J = 8.4 Hz, 1H), 6.53 (s, 2H), 3.55 (s, 2H), 3.38 (s,2H), 1.58 (d, J = 4.7 Hz, 2H), 1.56 (s, 2H), 1.52 (s, 2H).

[0082] Example 7

[0083] Except for replacing N,N-dimethylcarbamoyl chloride with 4-morpholinocarbamoyl chloride, the same procedure as in Example 1 was followed to prepare 3-(4-morpholinocarbamoyloxy)-4-hydroxybenzophenone (1OMQ) as a white solid with a yield of 68%.

[0084] mp: 146.7-147.3℃; ESI-MS: m / z 328.23 (M+1) + C 18 H 17 NO5 (327.3).

[0085] 1 H NMR (600 MHz, DMSO-d6) δ 10.72 (s, 1H), 7.68 – 7.66 (m, 2H), 7.66– 7.62 (m, 1H), 7.57 – 7.53 (m, 3H), 7.46 (d, J = 2.2 Hz, 1H), 7.04 (d, J =8.5 Hz, 1H), 3.64 (s, 4H), 3.58 (s, 2H), 3.40 (s, 2H).

[0086] Example 8

[0087] Except for replacing N,N-dimethylcarbamoyl chloride with 1-pyrrolidinecarbonyl chloride, the same procedure as in Example 1 was followed to prepare 3-(1-pyrrolidinecarbonyloxy)-4-hydroxybenzophenone (1OLQ) as a white solid with a yield of 70%.

[0088] mp: 154.1-156.8℃; ESI-MS: m / z 334.29 (M+NA) + C 18 H 17 NO4 (311.3).

[0089] 1H NMR (600 MHz, DMSO-d6) δ 10.64 (s, 1H), 7.68 – 7.66 (m, 2H), 7.64(t, J = 7.4 Hz, 1H), 7.55 (d, J = 7.6 Hz, 2H), 7.53 (s, 1H), 7.43 (d, J = 2.2Hz, 1H), 7.04 (d, J = 8.4 Hz, 1H), 3.50 (t, J = 6.7 Hz, 2H), 3.31 (t, J = 6.7Hz, 2H), 1.87 (dq, J = 22.7, 6.4 Hz, 4H).

[0090] Example 9

[0091] Except for replacing benzoic acid with p-methylbenzoic acid, the same procedure was followed as in Example 1 to prepare 4'-methyl-3-(N,N-dimethylformyloxy)-4-hydroxybenzophenone (JOJQ) as a white solid with a yield of 68%.

[0092] mp: 132.0-134.6℃; ESI-MS: m / z 300.95 (M+1) + C 17 H 17 NO4 (299.3).

[0093] 1 H NMR (600 MHz, DMSO-d6) δ 10.62 (s, 1H), 7.59 (d, J = 8.0 Hz, 2H), 7.50 (dd, J = 8.4, 2.2 Hz, 1H), 7.39 (d, J = 2.2 Hz, 1H), 7.34 (d, J = 7.9Hz, 2H), 7.03 (d, J = 8.4 Hz, 1H), 3.04 (s, 3H), 2.89 (s, 3H), 2.39 (s, 3H).

[0094] Example 10

[0095] Except for replacing benzoic acid with p-methylbenzoic acid, the same procedure was followed as in Example 2 to prepare 4'-methyl-3-(N,N-diethylcarbamoyloxy)-4-hydroxybenzophenone (JOYQ) as a white solid with a yield of 60%.

[0096] mp: 81.8-88.9℃; ESI-MS: m / z 328.22 (M+1) + C 19 H 21 NO4 (327.4).

[0097] 1 H NMR (600 MHz, DMSO-d6) δ 10.65 (s, 1H), 7.59 (d, J = 8.0 Hz, 2H), 7.49 (dd, J = 8.4, 2.2 Hz, 1H), 7.40 (d, J = 2.1 Hz, 1H), 7.35 (d, J = 7.9Hz, 2H), 7.02 (d, J = 8.5 Hz, 1H), 3.42 – 3.39 (m, 2H), 3.28 (q, J = 7.1 Hz,2H), 2.39 (s, 3H), 1.20 (t, J = 7.1 Hz, 3H), 1.10 (t, J = 7.0 Hz, 3H).

[0098] Example 11

[0099] Except for replacing benzoic acid with p-methylbenzoic acid, the same procedure was followed as in Example 3 to prepare 4'-methyl-3-(N-ethyl-N-methylcarbamoyloxy)-4-hydroxybenzophenone (JONQ) as a white solid with a yield of 50%.

[0100] mp: 125.9-128.7℃; ESI-MS: m / z 314.24 (M+1) + C 18 H 19 NO4 (313.4).

[0101] 1 H NMR (600 MHz, DMSO-d6) δ 10.65 (s, 1H), 7.60 (d, J = 7.9 Hz, 2H), 7.50 (dd, J= 8.4, 2.2 Hz, 1H), 7.40 (d, J = 2.1 Hz, 1H), 7.35 (d, J = 7.9Hz, 2H), 7.03 (d, J = 8.4 Hz, 1H), 3.43 (q, J = 7.0 Hz, 1H), 3.29 (q, J = 7.2Hz, 1H), 2.95 (d, J = 82.6 Hz, 3H), 2.40 (s, 3H), 1.14 (dt, J = 60.4, 7.2 Hz, 3H).

[0102] Example 12

[0103] Except for replacing benzoic acid with p-methylbenzoic acid, the same procedure was followed as in Example 4 to prepare 4'-methyl-3-(N,N-diisopropylcarbamoyloxy)-4-hydroxybenzophenone (JOIQ) as a white solid with a yield of 54%.

[0104] mp: 178.5-180.9℃; ESI-MS: m / z 356.80 (M+1) + C 21 H 25 NO4 (355.4).

[0105] 1 H NMR (600 MHz, DMSO-d6) δ 10.64 (s, 1H), 7.60 (d, J = 7.8 Hz, 2H), 7.48 (dd, J = 8.4, 2.2 Hz, 1H), 7.40 (d, J = 2.2 Hz, 1H), 7.36 (d, J = 7.8Hz, 2H), 7.02 (d, J = 8.4 Hz, 1H), 3.99 (d, J = 30.6 Hz, 2H), 2.40 (s, 3H), 1.25 (d, J = 35.5 Hz, 12H).

[0106] Example 13

[0107] Except for replacing benzoic acid with p-methylbenzoic acid, the same procedure was followed as in Example 5 to prepare 4'-methyl-3-[N,N-di(2-chloroethyl)carbamoyloxy]-4-hydroxybenzophenone (JOCQ) as a white solid with a yield of 55%.

[0108] mp: 118.6-120.0℃; ESI-MS: m / z 397.65 (M+1) + C 19 H 19 C l2 NO4 (396.3).

[0109] 1 H NMR (600 MHz, DMSO-d6) δ 10.80 (s, 1H), 7.60 (d, J = 7.7 Hz, 2H), 7.52 (d, J = 8.3 Hz, 1H), 7.50 (s, 1H), 7.36 (d, J = 8.0 Hz, 2H), 7.04 (d, J = 8.4 Hz, 1H), 3.93 (t, J = 6.5 Hz, 2H), 3.82 – 3.77 (m, 4H), 3.68 (t, J =6.5 Hz, 2H), 2.41 (d, J = 5.2 Hz, 3H).

[0110] Example 14

[0111] Except for replacing benzoic acid with p-methylbenzoic acid, the same procedure was followed as in Example 6 to prepare 4'-methyl-3-(1-piperidinylcarbonyloxy)-4-hydroxybenzophenone (JOPQ) as a white solid with a yield of 63%.

[0112] mp: 86.6-87.3℃; ESI-MS: m / z 339.33 (M), C 20 H 21 NO4 (339.4).

[0113] 1 H NMR (600 MHz, DMSO-d6) δ 10.64 (s, 1H), 7.59 (d, J = 8.0 Hz, 2H), 7.49 (dd, J = 8.4, 2.2 Hz, 1H), 7.39 (d, J= 2.2 Hz, 1H), 7.35 (d, J = 8.0Hz, 2H), 7.02 (d, J = 8.5 Hz, 1H), 3.55 (s, 2H), 3.45 (s, 2H), 2.40 (d, J =4.9 Hz, 3H), 1.58 (d, J = 4.9 Hz, 2H), 1.56 (s, 2H), 1.52 (s, 2H).

[0114] Example 15

[0115] Except for replacing benzoic acid with p-methylbenzoic acid, the same procedure was followed as in Example 7 to prepare 4'-methyl-3-(4-morpholinocarbonyloxy)-4-hydroxybenzophenone (JOMQ) as a white solid with a yield of 67%.

[0116] mp: 152.7-155.9℃; ESI-MS: m / z 340.17 (M-1) - C 19 H 19 NO5 (341.4).

[0117] 1 H NMR (600 MHz, DMSO-d6) δ 11.00 – 10.35 (m, 1H), 7.60 (d, J = 7.9Hz, 2H), 7.52 (dd, J = 8.4, 2.2 Hz, 1H), 7.44 (d, J = 2.1 Hz, 1H), 7.35 (d, J = 7.9 Hz, 2H), 7.04 (d, J = 8.5 Hz, 1H), 3.65 (s, 4H), 3.59 (s, 2H), 3.40 (s,2H), 2.41 (d, J = 4.9 Hz, 3H).

[0118] Example 16

[0119] Except for replacing benzoic acid with p-methylbenzoic acid, the same procedure was followed as in Example 8 to prepare 4'-methyl-3-(1-pyrrolidinecarbonyloxy)-4-hydroxybenzophenone (JOLQ) as a white solid with a yield of 70%.

[0120] mp: 136.2-138.5℃; ESI-MS: m / z 326.24 (M+1) + C 19 H 19 NO4 (325.4).

[0121] 1 H NMR (600 MHz, DMSO-d6) δ 10.59 (s, 1H), 7.59 (d, J = 8.0 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.41 (d, J = 2.2 Hz, 1H), 7.35 (d, J = 7.9Hz, 2H), 7.04 (d, J = 8.5 Hz, 1H), 3.51 (t, J = 6.7 Hz, 2H), 3.32 (t, J = 6.7Hz, 2H), 2.40 (s, 3H), 1.87 (dp, J = 25.6, 6.5 Hz, 4H).

[0122] Example 17

[0123] Except for replacing benzoic acid with p-ethylbenzoic acid, the same procedure was followed as in Example 1 to prepare 4'-ethyl-3-(N,N-dimethylcarbamoyloxy)-4-hydroxybenzophenone (2OJQ) as a white solid with a yield of 68%.

[0124] mp: 131.7-133.0℃; ESI-MS: m / z 314.24 (M+1) + C 18 H 19 NO4 (313.4).

[0125] 1 H NMR (600 MHz, DMSO-d6) δ 10.62 (s, 1H), 7.61 (d, J = 8.1 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.40 (d, J = 2.2 Hz, 1H), 7.38 (d, J = 7.9Hz, 2H), 7.03 (d, J= 8.4 Hz, 1H), 3.04 (s, 3H), 2.89 (s, 3H), 2.69 (q, J =7.6 Hz, 2H), 1.22 (t, J = 7.6 Hz, 3H).

[0126] Example 18

[0127] Except for replacing benzoic acid with p-ethylbenzoic acid, the same procedure was followed as in Example 2 to prepare 4'-ethyl-3-(N,N-diethylcarbamoyloxy)-4-hydroxybenzophenone (2OYQ) as a white solid with a yield of 60%.

[0128] mp: 79.2-81.2℃; ESI-MS: m / z 364.38 (M+NA) + C 20 H 23 NO4 (341.4).

[0129] 1 H NMR (600 MHz, DMSO-d6) δ 10.66 (s, 1H), 7.62 (d, J = 8.0 Hz, 2H), 7.50 (dd, J = 8.4, 2.2 Hz, 1H), 7.41 (d, J = 2.1 Hz, 1H), 7.39 (d, J = 7.9Hz, 2H), 7.03 (d, J = 8.4 Hz, 1H), 3.44 – 3.40 (m, 2H), 3.28 (t, J = 7.2 Hz, 2H), 2.70 (q, J = 7.6 Hz, 2H), 1.25 – 1.19 (m, 6H), 1.11 (t, J = 7.1 Hz, 3H).

[0130] Example 19

[0131] Except for replacing benzoic acid with p-ethylbenzoic acid, the same procedure was followed as in Example 3 to prepare 4'-ethyl-3-(N-ethyl-N-methylcarbamoyloxy)-4-hydroxybenzophenone (2ONQ) as a white solid with a yield of 66%.

[0132] mp: 136.8-139.9℃; ESI-MS: m / z 328.29 (M+1) +C 19 H 21 NO4 (327.4).

[0133] 1 H NMR (600 MHz, DMSO-d6) δ 10.65 (d, J = 17.7 Hz, 1H), 7.62 (d, J =8.1 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.41 (d, J = 2.2 Hz, 1H), 7.39(d, J = 7.9 Hz, 2H), 7.03 (d, J = 8.4 Hz, 1H), 3.43 (q, J = 7.0 Hz, 1H), 3.30(q, J = 7.1 Hz, 1H), 2.96 (d, J = 82.5 Hz, 3H), 2.70 (q, J = 7.6 Hz, 2H), 1.23 (t, J = 7.6 Hz, 3H), 1.14 (dt, J = 60.9, 7.1 Hz, 3H).

[0134] Example 20

[0135] Except for replacing benzoic acid with p-ethylbenzoic acid, the same procedure was followed as in Example 4 to prepare 4'-ethyl-3-(N,N-diisopropylcarbamoyloxy)-4-hydroxybenzophenone (2OIQ) as a white solid with a yield of 59%.

[0136] mp: 136.2-137.5℃; ESI-MS: m / z 370.11 (M+1) + C 22 H 27 NO4 (369.5).

[0137] 1 H NMR (600 MHz, DMSO-d6) δ 10.63 (s, 1H), 7.63 (d, J = 8.1 Hz, 2H), 7.48 (dd, J = 8.4, 2.2 Hz, 1H), 7.40 (d, J= 2.1 Hz, 1H), 7.39 (d, J = 8.0Hz, 2H), 7.02 (d, J = 8.4 Hz, 1H), 3.99 (d, J = 32.0 Hz, 2H), 2.70 (q, J =7.6 Hz, 2H), 1.34 – 1.17 (m, 15H).

[0138] Example 21

[0139] Except for replacing benzoic acid with p-ethylbenzoic acid, the same procedure was followed as in Example 5 to prepare 4'-ethyl-3-[N,N-di(2-chloroethyl)carbamoyloxy]-4-hydroxybenzophenone (2OCQ) as a white solid with a yield of 51%.

[0140] mp: 130.9-132.3℃; ESI-MS: m / z 411.40 (M+1) + C 20 H 21 C l2 NO4 (410.3).

[0141] 1 H NMR (600 MHz, DMSO-d6) δ 10.79 (s, 1H), 7.63 (d, J = 7.9 Hz, 2H), 7.53 (dd, J = 8.4, 2.2 Hz, 1H), 7.50 (d, J = 2.2 Hz, 1H), 7.39 (d, J = 8.0Hz, 2H), 7.04 (d, J = 8.4 Hz, 1H), 3.93 (t, J = 6.5 Hz, 2H), 3.79 (dt, J =10.0, 6.5 Hz, 4H), 3.67 (t, J = 6.5 Hz, 2H), 2.70 (q, J = 7.6 Hz, 2H), 1.23(t, J = 7.6 Hz, 3H).

[0142] Example 22

[0143] Except for replacing benzoic acid with p-ethylbenzoic acid, the same procedure was followed as in Example 6 to prepare 4'-ethyl-3-(1-piperidinylcarbonyloxy)-4-hydroxybenzophenone (2OPQ) as a white solid with a yield of 60%.

[0144] mp: 137.2-139.4℃; ESI-MS: m / z 354.33 (M+1) + C 21 H 23 NO4 (353.4).

[0145] 1 H NMR (600 MHz, DMSO-d6) δ 10.65 (s, 1H), 7.62 (d, J = 8.0 Hz, 2H), 7.50 (dd, J = 8.4, 2.2 Hz, 1H), 7.41 (d, J = 2.1 Hz, 1H), 7.39 (d, J = 7.9Hz, 2H), 7.03 (d, J = 8.4 Hz, 1H), 3.55 (s, 2H), 3.48 (s, 2H), 2.70 (q, J =7.6 Hz, 2H), 1.60 – 1.58 (m, 2H), 1.57 (s, 2H), 1.52 (s, 2H), 1.23 (t, J =7.6 Hz, 3H).

[0146] Example 23

[0147] Except for replacing benzoic acid with p-ethylbenzoic acid, the same procedure was followed as in Example 7 to prepare 4'-ethyl-3-(4-morpholinocarbonyloxy)-4-hydroxybenzophenone (2OMQ) as a white solid with a yield of 65%.

[0148] mp: 149.2-152.8℃; ESI-MS: m / z356 (M+1) + C 20 H 21 NO5 (355.4).

[0149] 1 H NMR (600 MHz, DMSO-d6) δ 10.67 (s, 1H), 7.62 (d, J = 8.2 Hz, 2H), 7.52 (dd, J= 8.4, 2.2 Hz, 1H), 7.44 (d, J = 2.2 Hz, 1H), 7.38 (d, J = 8.1Hz, 2H), 7.04 (d, J = 8.4 Hz, 1H), 3.64 (s, 4H), 3.58 (s, 2H), 3.40 (s, 2H), 2.70 (q, J = 7.6 Hz, 2H), 1.22 (t, J = 7.6 Hz, 3H).

[0150] Example 24

[0151] Except for replacing benzoic acid with p-ethylbenzoic acid, the same procedure was followed as in Example 8 to prepare 4'-ethyl-3-(1-pyrrolidinecarbonyloxy)-4-hydroxybenzophenone (2OLQ) as a white solid with a yield of 69%.

[0152] mp: 151-153.8℃; ESI-MS: m / z 339.62 (M) + C 20 H 21 NO4 (339.4).

[0153] 1 H NMR (600 MHz, DMSO-d6) δ 10.59 (s, 1H), 7.62 (d, J = 7.9 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.42 (d, J = 2.2 Hz, 1H), 7.39 (d, J = 7.9Hz, 2H), 7.04 (d, J = 8.4 Hz, 1H), 3.51 (t, J = 6.7 Hz, 2H), 3.32 (t, J = 6.7Hz, 2H), 2.70 (q, J = 7.6 Hz, 2H), 1.87 (dp, J = 25.8, 6.6 Hz, 4H), 1.23 (t, J = 7.6 Hz, 3H).

[0154] Example 25

[0155] Except for replacing benzoic acid with p-propylbenzoic acid, the same procedure was followed as in Example 1 to prepare 4'-propyl-3-(N,N-dimethylcarbamoyloxy)-4-hydroxybenzophenone (3OJQ) as a white solid with a yield of 60%.

[0156] mp: 101.9-102.1℃; ESI-MS: m / z 350.19 (M+NA) + C 19 H 21 NO4 (327.4).

[0157] 1 H NMR (600 MHz, DMSO-d6) δ 10.63 (s, 1H), 7.61 (d, J = 8.1 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.41 (d, J = 2.2 Hz, 1H), 7.36 (d, J = 8.0Hz, 2H), 7.04 (d, J = 8.4 Hz, 1H), 3.05 (d, J = 9.4 Hz, 3H), 2.91 (d, J =10.3 Hz, 3H), 2.65 (t, J = 7.6 Hz, 2H), 1.64 (h, J = 7.3 Hz, 2H), 0.92 (t, J = 7.3 Hz, 3H).

[0158] Example 26

[0159] Except for replacing benzoic acid with p-propylbenzoic acid, the same procedure was followed as in Example 2 to prepare 4'-propyl-3-(N,N-diethylcarbamoyloxy)-4-hydroxybenzophenone (3OYQ) as a white solid with a yield of 57%.

[0160] mp: 78.8-80.3℃; ESI-MS: m / z 378.30 (M+NA) + C 21 H 25 NO4 (355.4).

[0161] 1 H NMR (600 MHz, DMSO-d6) δ 10.66 (s, 1H), 7.62 (d,J = 8.1 Hz, 2H), 7.50 (dd, J = 8.4, 2.2 Hz, 1H), 7.41 (d, J = 2.2 Hz, 1H), 7.37 (d, J = 7.9Hz, 2H), 7.03 (d, J = 8.4 Hz, 1H), 3.44 – 3.40 (m, 2H), 3.29 (q, J = 7.0 Hz, 2H), 2.65 (t, J = 7.6 Hz, 2H), 1.64 (h, J = 7.4 Hz, 2H), 1.16 (dt, J = 63.1, 7.0 Hz, 6H), 0.92 (t, J = 7.3 Hz, 3H).

[0162] Example 27

[0163] Except for replacing benzoic acid with p-propylbenzoic acid, the same procedure was followed as in Example 3 to prepare 4'-propyl-3-(N-ethyl-N-methylcarbamoyloxy)-4-hydroxybenzophenone (3ONQ) as a white solid with a yield of 68%.

[0164] mp: 92.9-94.5℃; ESI-MS: m / z 342.32 (M+1) + C 20 H 23 NO4 (341.4).

[0165] 1 H NMR (600 MHz, DMSO-d6) δ 10.65 (s, 1H), 7.62 (d, J = 8.1 Hz, 2H), 7.50 (dd, J = 8.4, 2.2 Hz, 1H), 7.41 (d, J = 2.2 Hz, 1H), 7.36 (d, J = 8.0Hz, 2H), 7.03 (d, J = 8.4 Hz, 1H), 3.43 (q, J = 7.0 Hz, 1H), 3.30 (q, J = 7.2Hz, 1H), 2.96 (d,J = 82.7 Hz, 3H), 2.65 (t, J = 7.6 Hz, 2H), 1.64 (h, J =7.4 Hz, 2H), 1.14 (dt, J = 60.7, 7.2 Hz, 3H), 0.92 (t, J = 7.3 Hz, 3H).

[0166] Example 28

[0167] Except for replacing benzoic acid with p-propylbenzoic acid, the same procedure was followed as in Example 4 to prepare 4'-propyl-3-(N,N-diisopropylcarbamoyloxy)-4-hydroxybenzophenone (3OIQ) as a white solid with a yield of 55%.

[0168] mp: 102.9-104.1℃; ESI-MS: m / z 384.2 (M+1) + C 23 H 29 NO4 (383.5).

[0169] 1 H NMR (600 MHz, DMSO-d6) δ 8.46 (s, 1H), 7.89 – 7.83 (m, 2H), 7.59(dd, J = 9.0, 2.0 Hz, 1H), 7.52 (d, J = 2.1 Hz, 1H), 7.37 – 7.30 (m, 2H), 7.02 (d, J = 8.9 Hz, 1H), 3.88 (hept, J = 6.5 Hz, 2H), 2.61 (tt, J = 6.3, 1.0Hz, 2H), 1.64 (dtd, J = 13.9, 7.5, 6.3 Hz, 2H), 1.20 (d, J = 6.4 Hz, 12H), 0.95 (t, J = 7.5 Hz, 3H).

[0170] Example 29

[0171] Except for replacing benzoic acid with p-propylbenzoic acid, the same procedure was followed as in Example 5 to prepare 4'-propyl-3-[N,N-di(2-chloroethyl)carbamoyloxy]-4-hydroxybenzophenone (3OCQ) as a white solid with a yield of 67%.

[0172] mp: 127.9-128.4℃; ESI-MS: m / z 425.7 (M+1) + C 21 H 23 C l2 NO4 (424.3).

[0173] 1 H NMR (600 MHz, DMSO-d6) δ 8.46 (s, 1H), 7.89 – 7.83 (m, 2H), 7.59(dd, J = 9.0, 2.0 Hz, 1H), 7.52 (d, J = 2.1 Hz, 1H), 7.37 – 7.30 (m, 2H), 7.02 (d, J = 8.9 Hz, 1H), 3.67 (t, J = 3.5 Hz, 4H), 3.56 (t, J = 3.5 Hz, 4H), 2.61 (tt, J = 6.3, 1.0 Hz, 2H), 1.64 (dtd, J = 13.9, 7.5, 6.3 Hz, 2H), 0.95(t, J = 7.5 Hz, 3H).

[0174] Example 30

[0175] Except for replacing benzoic acid with p-propylbenzoic acid, the same procedure was followed as in Example 6 to prepare 4'-propyl-3-(1-piperidinylcarbonyloxy)-4-hydroxybenzophenone (3OPQ) as a white solid with a yield of 56%.

[0176] mp: 136.6-138.4℃; ESI-MS: m / z 390.00 (M+NA) + C 22 H 25 NO4 (367.4).

[0177] 1H NMR (600 MHz, DMSO-d6) δ 10.65 (s, 1H), 7.63 – 7.60 (m, 2H), 7.50(dd, J = 8.4, 2.2 Hz, 1H), 7.41 (d, J = 2.1 Hz, 1H), 7.36 (d, J = 8.0 Hz, 2H), 7.03 (d, J = 8.4 Hz, 1H), 3.56 (s, 3H), 2.65 (t, J = 7.6 Hz, 2H), 1.64(d, J = 7.5 Hz, 2H), 1.59 (d, J = 4.9 Hz, 2H), 1.56 (s, 1H), 1.52 (s, 2H), 0.92 (t, J = 7.3 Hz, 4H).

[0178] Example 31

[0179] Except for replacing benzoic acid with p-propylbenzoic acid, the same procedure was followed as in Example 7 to prepare 4'-propyl-3-(4-morpholinocarbonyloxy)-4-hydroxybenzophenone (3OMQ) as a white solid with a yield of 68%.

[0180] mp: 128.2-129.5℃; ESI-MS: m / z 368.38 (M-1) - C 21 H 23 NO5 (369.4).

[0181] 1 H NMR (600 MHz, DMSO-d6) δ 10.68 (s, 1H), 7.62 (d, J = 8.1 Hz, 2H), 7.52 (dd, J = 8.4, 2.2 Hz, 1H), 7.45 (d, J = 2.2 Hz, 1H), 7.36 (d, J = 8.0Hz, 2H), 7.04 (d, J = 8.4 Hz, 1H), 3.65 (s, 6H), 2.65 (t, J = 7.6 Hz, 2H), 1.64 (h, J= 7.4 Hz, 3H), 0.92 (t, J = 7.3 Hz, 4H).

[0182] Example 32

[0183] Except for replacing benzoic acid with p-propylbenzoic acid, the same procedure was followed as in Example 8 to prepare 4'-propyl-3-(1-pyrrolidinecarbonyloxy)-4-hydroxybenzophenone (3OLQ) as a white solid with a yield of 70%.

[0184] mp: 125.6-126.3℃; ESI-MS: m / z 354.30 (M+1) + C 21 H 23 NO4 (353.4).

[0185] 1 H NMR (600 MHz, DMSO-d6) δ 10.59 (s, 1H), 7.61 (d, J = 8.1 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.42 (d, J = 2.2 Hz, 1H), 7.36 (d, J = 8.0Hz, 2H), 7.04 (d, J = 8.5 Hz, 1H), 3.51 (t, J = 6.7 Hz, 2H), 3.32 (t, J = 6.7Hz, 2H), 2.65 (t, J = 7.6 Hz, 2H), 1.87 (dp, J = 25.7, 6.5 Hz, 4H), 1.64 (h, J = 7.4 Hz, 2H), 0.92 (t, J = 7.3 Hz, 3H).

[0186] Example 33

[0187] Except for replacing benzoic acid with p-butylbenzoic acid, the same procedure was followed as in Example 1 to prepare 4'-butyl-3-(N,N-dimethylcarbamoyloxy)-4-hydroxybenzophenone (4OJQ) as a white solid with a yield of 61%.

[0188] mp: 93.9-94.7℃; ESI-MS: m / z 364.23 (M+NA)+ C 20 H 23 NO4 (341.4).

[0189] 1 H NMR (600 MHz, DMSO-d6) δ 10.61 (s, 1H), 7.61 (d, J = 8.1 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.40 (d, J = 2.2 Hz, 1H), 7.36 (d, J = 7.9Hz, 2H), 7.03 (d, J = 8.5 Hz, 1H), 3.05 (d, J = 9.4 Hz, 3H), 2.91 (d, J =10.3 Hz, 3H), 2.67 (t, J = 7.7 Hz, 2H), 1.60 (p, J = 7.6 Hz, 2H), 1.33 (h, J = 7.4 Hz, 2H), 0.91 (t, J = 7.4 Hz, 3H).

[0190] Example 34

[0191] Except for replacing benzoic acid with p-butylbenzoic acid, the same procedure was followed as in Example 2 to prepare 4'-butyl-3-(N,N-diethylcarbamoyloxy)-4-hydroxybenzophenone (4OYQ) as a white solid with a yield of 76%.

[0192] mp: 87.7-89.2℃; ESI-MS: m / z 370.35 (M+1) + C 22 H 27 NO4 (369.5).

[0193] 1 H NMR (600 MHz, DMSO-d6) δ 10.63 (s, 1H), 7.61 (d, J = 7.9 Hz, 2H), 7.49 (dd, J = 8.4, 2.2 Hz, 1H), 7.40 (d, J = 2.2 Hz, 1H), 7.37 (d,J = 7.9Hz, 2H), 7.02 (d, J = 8.4 Hz, 1H), 3.41 (d, J = 7.0 Hz, 2H), 3.28 (d, J = 7.3Hz, 2H), 2.67 (t, J = 7.7 Hz, 2H), 1.60 (p, J = 7.6 Hz, 2H), 1.33 (h, J = 7.4Hz, 2H), 1.21 (t, J = 7.2 Hz, 3H), 1.10 (t, J = 7.2 Hz, 3H), 0.91 (t, J = 7.4Hz, 3H).

[0194] Example 35

[0195] Except for replacing benzoic acid with p-butylbenzoic acid, the same procedure was followed as in Example 3 to prepare 4'-butyl-3-(N-ethyl-N-methylcarbamoyloxy)-4-hydroxybenzophenone (4ONQ) as a white solid with a yield of 67%.

[0196] mp: 55.2-57.5℃; ESI-MS: m / z 356.31 (M+1) + C 21 H 25 NO4 (355.4).

[0197] 1 H NMR (600 MHz, DMSO-d6) δ 10.63 (s, 1H), 7.61 (d, J = 8.1 Hz, 2H), 7.50 (dd, J = 8.4, 2.2 Hz, 1H), 7.41 (d, J = 2.2 Hz, 1H), 7.36 (d, J = 7.9Hz, 2H), 7.03 (d, J = 8.4 Hz, 1H), 3.43 (q, J = 6.9 Hz, 1H), 3.29 (q, J =7.2, 6.6 Hz, 1H), 2.96 (dd, J= 82.6, 8.5 Hz, 3H), 2.67 (t, J = 7.7 Hz, 2H), 1.60 (p, J = 7.5 Hz, 2H), 1.33 (h, J = 7.4 Hz, 2H), 1.15 (dq, J = 60.7, 7.6Hz, 3H), 0.91 (t, J = 7.4 Hz, 3H).

[0198] Example 36

[0199] Except for replacing benzoic acid with p-butylbenzoic acid, the same procedure was followed as in Example 4 to prepare 4'-butyl-3-(N,N-diisopropylcarbamoyloxy)-4-hydroxybenzophenone (4OIQ) as a white solid with a yield of 78%.

[0200] mp: 136.7-137.1℃; ESI-MS: m / z 398.1 (M+1) + C 24 H 31 NO4 (397.5).

[0201] 1 H NMR (600 MHz, DMSO-d6) δ 8.46 (s, 1H), 7.95 – 7.75 (m, 2H), 7.59(dd, J = 9.0, 2.0 Hz, 1H), 7.52 (d, J = 2.1 Hz, 1H), 7.40 – 7.35 (m, 2H), 7.02 (d, J = 8.9 Hz, 1H), 3.88 (hept, J = 6.5 Hz, 2H), 2.62 (tt, J = 8.1, 1.0Hz, 2H), 1.62 – 1.53 (m, 2H), 1.35 (hept, J = 6.7 Hz, 2H), 1.20 (d, J = 6.4Hz, 12H), 0.94 (t, J = 7.2 Hz, 3H).

[0202] Example 37

[0203] Except for replacing benzoic acid with p-butylbenzoic acid, the same procedure was followed as in Example 5 to prepare 4'-butyl-3-[N,N-di(2-chloroethyl)carbamoyloxy]-4-hydroxybenzophenone (4OCQ) as a white solid with a yield of 72%.

[0204] mp: 128.2-130.1℃; ESI-MS: m / z 439.7 (M+1) + C 22 H 25 C l2 NO4 (438.3).

[0205] 1 H NMR (600 MHz, DMSO-d6) δ 8.46 (s, 1H), 7.95 – 7.75 (m, 2H), 7.67 –7.47 (m, 2H), 7.41 – 7.34 (m, 2H), 7.02 (d, J = 8.9 Hz, 1H), 3.67 (t, J = 3.5Hz, 4H), 3.56 (t, J = 3.5 Hz, 4H), 2.62 (tt, J = 8.2, 1.0 Hz, 2H), 1.63 –1.53 (m, 2H), 1.34 (h, J = 7.1 Hz, 2H), 0.94 (t, J = 7.2 Hz, 3H).

[0206] Example 38

[0207] Except for replacing benzoic acid with p-butylbenzoic acid, the same procedure was followed as in Example 6 to prepare 4'-butyl-3-(1-piperidinylcarbonyloxy)-4-hydroxybenzophenone (4OPQ) as a white solid with a yield of 71%.

[0208] mp: 121.6-123.8℃; ESI-MS: m / z 382.36 (M+1) + C 23 H 27 NO4 (381.5).

[0209] 1 H NMR (600 MHz, DMSO-d6) δ 10.63 (s, 1H), 7.61 (d, J = 7.9 Hz, 2H), 7.50 (dd, J= 8.4, 2.2 Hz, 1H), 7.41 (d, J = 2.2 Hz, 1H), 7.36 (d, J = 7.9Hz, 2H), 7.03 (d, J = 8.4 Hz, 1H), 3.56 (s, 2H), 3.38 (s, 2H), 2.67 (t, J =7.6 Hz, 2H), 1.59 (q, J = 7.9 Hz, 6H), 1.53 (s, 2H), 1.33 (h, J = 7.4 Hz, 2H), 0.91 (t, J = 7.4 Hz, 3H).

[0210] Example 39

[0211] Except for replacing benzoic acid with p-butylbenzoic acid, the same procedure was followed as in Example 7 to prepare 4'-butyl-3-(4-morpholinocarbonyloxy)-4-hydroxybenzophenone (4OMQ) as a white solid with a yield of 77%.

[0212] mp: 118.7-120.8℃; ESI-MS: m / z 382.45 (M-1) - C 22 H 25 NO5 (383.4).

[0213] 1 H NMR (600 MHz, DMSO-d6) δ 10.65 (s, 1H), 7.61 (d, J = 7.9 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.44 (d, J = 2.1 Hz, 1H), 7.36 (d, J = 8.0Hz, 2H), 7.04 (d, J = 8.4 Hz, 1H), 3.64 (s, 4H), 3.58 (s, 2H), 3.40 (s, 2H), 2.67 (t, J = 7.6 Hz, 2H), 1.59 (p, J = 7.5 Hz, 2H), 1.33 (h, J = 7.4 Hz, 2H), 0.91 (t,J = 7.4 Hz, 3H).

[0214] Example 40

[0215] Except for replacing benzoic acid with p-butylbenzoic acid, the same procedure was followed as in Example 8 to prepare 4'-butyl-3-(1-pyrrolidinecarbonyloxy)-4-hydroxybenzophenone (4OLQ) as a white solid with a yield of 74%.

[0216] mp: 135.2-136.9℃; ESI-MS: m / z 366.62 (M+1) + C 22 H 25 NO4 (367.4).

[0217] 1 H NMR (600 MHz, DMSO-d6) δ 10.57 (s, 1H), 7.61 (d, J = 8.1 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.42 (d, J = 2.2 Hz, 1H), 7.36 (d, J = 7.9Hz, 2H), 7.04 (d, J = 8.4 Hz, 1H), 3.51 (t, J = 6.7 Hz, 2H), 3.32 (t, J = 6.7Hz, 2H), 2.67 (t, J = 7.7 Hz, 2H), 1.87 (dq, J = 22.7, 6.4 Hz, 4H), 1.60 (p, J = 7.5 Hz, 2H), 1.33 (h, J = 7.4 Hz, 2H), 0.91 (t, J = 7.3 Hz, 3H).

[0218] Example 41

[0219] Except for replacing benzoic acid with p-tert-butylbenzoic acid, the same procedure was followed as in Example 1 to prepare 4'-tert-butyl-3-(N,N-dimethylcarbamoyloxy)-4-hydroxybenzophenone (4IOJQ) as a white solid with a yield of 76%.

[0220] mp: 171.0-173.8℃; ESI-MS: m / z 342.91 (M+1) + C 20 H 23 NO4 (341.4).

[0221] 1 H NMR (600 MHz, DMSO-d6) δ 10.61 (s, 1H), 7.64 (d, J = 8.4 Hz, 2H),7.59 – 7.55 (m, 2H), 7.52 (dd, J = 8.4, 2.2 Hz, 1H), 7.42 (d, J = 2.2 Hz, 1H), 7.04 (d, J = 8.4 Hz, 1H), 3.05 (d, J = 9.1 Hz, 3H), 2.91 (d, J = 10.1Hz, 3H), 1.33 (s, 9H).

[0222] Example 42

[0223] Except for replacing benzoic acid with p-tert-butylbenzoic acid, the same procedure was followed as in Example 2 to prepare 4'-tert-butyl-3-(N,N-diethylcarbamoyloxy)-4-hydroxybenzophenone (4IOYQ) as a white solid with a yield of 68%.

[0224] mp: 87.2-89.1℃; ESI-MS: m / z 370.36 (M+1) + C 22 H 27 NO4 (369.5).

[0225] 1 H NMR (600 MHz, DMSO-d6) δ 10.65 (s, 1H), 7.65 (d, J = 8.4 Hz, 2H), 7.57 (d, J = 8.5 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.43 (d, J = 2.2Hz, 1H), 7.03 (d, J = 8.4 Hz, 1H), 3.41 (q, J= 7.1 Hz, 2H), 3.29 (q, J = 7.0Hz, 2H), 1.33 (s, 9H), 1.21 (t, J = 6.8 Hz, 3H), 1.11 (t, J = 7.0 Hz, 3H).

[0226] Example 43

[0227] Except for replacing benzoic acid with p-tert-butylbenzoic acid, the same procedure was followed as in Example 3 to prepare 4'-tert-butyl-3-(N-ethyl-N-methylcarbamoyloxy)-4-hydroxybenzophenone (4IONQ) as a white solid with a yield of 60%.

[0228] mp: 133.5-135.6℃; ESI-MS: m / z 356.46 (M+1) + C 21 H 25 NO4 (355.4).

[0229] 1 H NMR (600 MHz, DMSO-d6) δ 10.63 (s, 1H), 7.64 (d, J = 8.5 Hz, 2H), 7.57 (d, J = 8.5 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.42 (d, J = 2.2Hz, 1H), 7.03 (d, J = 8.4 Hz, 1H), 3.43 (q, J = 6.9 Hz, 1H), 3.30 (q, J = 7.3Hz, 1H), 2.96 (d, J = 82.5 Hz, 3H), 1.33 (s, 9H), 1.14 (dt, J = 61.6, 7.1 Hz, 3H).

[0230] Example 44

[0231] Except for replacing benzoic acid with p-tert-butylbenzoic acid, the same procedure was followed as in Example 4 to prepare 4'-tert-butyl-3-(N,N-diisopropylcarbamoyloxy)-4-hydroxybenzophenone (4IOIQ) as a white solid with a yield of 55%.

[0232] mp: 97.6-100.8℃; ESI-MS: m / z 398.38 (M+1) + C 24 H 31 NO4 (397.5).

[0233] 1 H NMR (600 MHz, DMSO-d6) δ 10.62 (s, 1H), 7.65 (d, J = 8.3 Hz, 2H), 7.57 (d, J = 8.4 Hz, 2H), 7.49 (dd, J = 8.4, 2.2 Hz, 1H), 7.42 (d, J = 2.2Hz, 1H), 7.02 (d, J = 8.4 Hz, 1H), 3.99 (d, J = 36.9 Hz, 2H), 1.33 (s, 9H), 1.25 (d, J = 34.8 Hz, 12H).

[0234] Example 45

[0235] Except for replacing benzoic acid with p-tert-butylbenzoic acid, the same procedure was followed as in Example 5 to prepare 4'-tert-butyl-3-[N,N-di(2-chloroethyl)carbamoyloxy]-4-hydroxybenzophenone (4IOCQ) as a white solid with a yield of 68%.

[0236] mp: 127.2-129.0℃; ESI-MS: m / z 439.61 (M+1) + C 22 H 25 C l2 NO4 (438.3).

[0237] 1 H NMR (600 MHz, DMSO-d6) δ 10.63 (s, 1H), 7.64 (d, J = 8.3 Hz, 2H), 7.57 (d, J = 8.4 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.42 (d, J = 2.2Hz, 1H), 7.03 (d, J= 8.4 Hz, 1H), 3.43 (q, J = 7.0 Hz, 1H), 3.30 (q, J = 7.2Hz, 1H), 3.02 (s, 1H), 2.89 (s, 1H), 1.33 (s, 9H), 1.20 (t, J = 7.1 Hz, 2H), 1.10 (q, J = 7.7, 7.1 Hz, 2H).

[0238] Example 46

[0239] Except for replacing benzoic acid with p-tert-butylbenzoic acid, the same procedure was followed as in Example 6 to prepare 4'-tert-butyl-3-(1-piperidinylcarbonyloxy)-4-hydroxybenzophenone (4IOPQ) as a white solid with a yield of 60%.

[0240] mp: 169.3-170.5℃; ESI-MS: m / z382.36 (M+1) + C 23 H 27 NO4 (381.5).

[0241] 1 H NMR (600 MHz, DMSO-d6) δ 10.63 (s, 1H), 7.64 (d, J = 8.3 Hz, 2H), 7.57 (d, J = 8.3 Hz, 2H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.42 (d, J = 2.2Hz, 1H), 7.03 (d, J = 8.4 Hz, 1H), 3.56 (s, 2H), 3.39 (s, 2H), 1.60 – 1.58(m, 2H), 1.56 (s, 2H), 1.53 (s, 2H), 1.33 (d, J = 1.9 Hz, 9H).

[0242] Example 47

[0243] Except for replacing benzoic acid with p-tert-butylbenzoic acid, the same procedure was followed as in Example 7 to prepare 4'-tert-butyl-3-(4-morpholinocarbonyloxy)-4-hydroxybenzophenone (4IOMQ) as a white solid with a yield of 60%.

[0244] mp: 179.5-182.1℃; ESI-MS: m / z382.33 (M-1) - C 22 H 25 NO5 (383.4).

[0245] 1 H NMR (600 MHz, DMSO-d6) δ 10.66 (s, 1H), 7.64 (d, J = 8.4 Hz, 2H), 7.57 (d, J = 8.5 Hz, 2H), 7.53 (dd, J = 8.4, 2.2 Hz, 1H), 7.46 (d, J = 2.2Hz, 1H), 7.04 (d, J = 8.4 Hz, 1H), 3.65 (s, 4H), 3.59 (s, 2H), 3.40 (s, 2H), 1.33 (d, J = 1.7 Hz, 9H).

[0246] Example 48

[0247] Except for replacing benzoic acid with p-tert-butylbenzoic acid, the same procedure was followed as in Example 8 to prepare 4'-tert-butyl-3-(1-pyrrolidinecarbonyloxy)-4-hydroxybenzophenone (4IOLQ) as a white solid with a yield of 79%.

[0248] mp: 175.1-176.5℃; ESI-MS: m / z368.87 (M+1) + C 22 H 25 NO4 (367.4).

[0249] 1 H NMR (600 MHz, DMSO-d6) δ 10.58 (s, 1H), 7.64 (d, J = 8.4 Hz, 2H), 7.57 (d, J = 8.4 Hz, 2H), 7.52 (dd, J = 8.5, 2.2 Hz, 1H), 7.43 (d, J = 2.2Hz, 1H), 7.04 (d, J = 8.4 Hz, 1H), 3.51 (t, J= 6.6 Hz, 2H), 3.32 (t, J = 6.7Hz, 2H), 1.87 (dq, J = 22.7, 6.4 Hz, 4H), 1.33 (s, 9H).

[0250] Example 49

[0251] In this embodiment, donepezil was used as a positive control to determine the inhibitory activities of acetylcholinesterase and butyrylcholinesterase in some typical compounds prepared in Examples 1 to 48.

[0252] Acetylcholinesterase ( ee AChE), butyrylcholinesterase ( eq BuChE), iodothioacetylcholine, iodothiobutyrylcholine and 5,5-dithiobis(2-nitrobenzoic acid) (DTNB) were all purchased from Sigma Aldrich.

[0253] Donepezil was purchased from MedChemExpress and used as a positive control.

[0254] Add 100 μL of buffer containing 5,5-dithiobis(2-nitrobenzoic acid) and 20 μL of different concentrations of the test compound or blank control to each well of a 96-well plate. Then add 50 μL of acetylcholinesterase or butyrylcholinesterase (0.22 U / mL) to the sample wells and standard control wells, mix well, and incubate at 37°C for 10 min. Finally, add 10 μL of iodothioacetylcholine or iodothiobutyrylcholine solution to each well, mix well, and incubate at 37°C for 15 min.

[0255] The absorbance of the samples at 412 nm was measured using an ELISA reader, and the inhibition rate of each compound against acetylcholinesterase or butyrylcholinesterase was calculated. All experiments were performed in triplicate, with each well being independently repeated three times.

[0256] Inhibition rate (%) = [1 - ( A 样品 - A 空白 ) / ( A 标准 - A 空白 )]×100% A 样品 This indicates the absorbance value corresponding to the addition of the compound, cholinesterase, substrate, and chromogenic agent; A 标准 This indicates the absorbance value corresponding to the addition of cholinesterase, substrate, and chromogenic agent; A 空白This indicates the absorbance value corresponding to the addition of only the substrate and the colorimetric agent.

[0257] The test results are shown in Table 2.

[0258]

[0259] As can be seen from Table 2, the listed compounds have... eq BuChE all showed good inhibitory activity, but for ee The inhibitory activity of AChE was low (IC50). 50 >100 μM). Donepezil (positive control) had a selectivity index (SI) of 0.01, but compound 3OIQ had the lowest SI of 3.53, which is 353 times that of donepezil, and compound JOPQ had the highest SI of 160.91, which is 16091 times that of donepezil. Therefore, compared with donepezil, the typical compounds listed in the table all exhibit high selectivity for BuChE.

[0260] Among them, 7 compounds are effective against eq BuChE's inhibitory activity was significantly superior to donepezil. P <0.05), including JOPQ (IC 50 =0.62±0.02μM), 2OPQ (IC 50 =0.96±0.03μM), 3OPQ (IC 50 =1.07±0.02μM), 4IOPQ (IC 50 =1.12±0.11μM), 4OPQ (IC 50 =1.61±0.05μM), 1OPQ (IC 50 =2.68±0.14μM), 4OLQ (IC 50 =4.87±0.04μM).

[0261] Example 50

[0262] This embodiment uses JOPQ as an example to test the kinetic parameters of the prepared compound.

[0263] Add 100 μL of buffer containing 5,5-dithiobis(2-nitrobenzoic acid), 20 μL of JOPO solution of different concentrations or blank control, and 50 μL of butyrylcholinesterase (0.22 U / mL) to each well of a 96-well plate. Mix well and incubate at 37°C for 10 min. Finally, add 10 μL of iodothiobutyrylcholine and control the final concentrations to 0.028 mM, 0.056 mM, 0.111 mM, 0.222 mM, 0.444 mM, and 0.889 mM to initiate the enzymatic reaction. Mix well and incubate at 37°C for 15 min.

[0264] The absorbance of the sample at 412 nm was measured over time using a microplate reader for 15 minutes. All experiments were performed in triplicate, with each well being independently repeated three times.

[0265] For each combination of inhibitor and substrate concentrations, a linear regression was performed on the absorbance of the product (TNB) against the reaction time t using the time-scan curve. The slope of the resulting line represents the enzyme-catalyzed reaction rate under those conditions. V ).

[0266] For each fixed inhibitor concentration, at 1 / V Plot the graph using Lineweaver-Burk with 1 / [S] as the ordinate and 1 / [S] as the abscissa (where [S] is the final substrate concentration) to obtain a Lineweaver-Burk fitted line.

[0267] Plot the Lineweaver-Burk fitted lines at different inhibitor concentrations on the same graph, and the intercepts of the four lines on the Y-axis (i.e., 1 / V max The value increases with increasing inhibitor concentration, but the intersection point on the X-axis (i.e., -1 / ) K m The inhibitor concentration remains constant, which is characteristic of non-competitive inhibition. A quadratic plot of the slopes of the four straight lines against the inhibitor concentration is then performed. Figure 1 The absolute value of the intersection point of the straight line and the X-axis is the suppression constant. K i The specific experimental results are listed in Table 3.

[0268]

[0269] This invention conducted a BuChE binding kinetics study on the representative compound JOPQ. Table 3 shows the enzyme kinetic parameters, indicating that as the concentration of JOPQ increases, V max From 0.02848 mM·L -1 ·min -1 Decreased to 0.002506 mM·L -1 ·min -1 This indicates that JOPQ has a significant inhibitory effect on BuChE, and the inhibition constant is [missing information]. K i The value was 0.09 μM, indicating a strong affinity between the two.

[0270] K m The values ​​remained stable between 445.3 and 487.1 μM, without a significant trend. This kinetic behavior is consistent with the characteristics of non-competitive inhibition, i.e. K mThe value remains basically unchanged. V max The value decreases with increasing compound concentration. A Lineweaver-Burk double reciprocal plot shows that the fitted lines at each concentration intersect at a point on the X-axis (-1 / ). K m This further confirms the mechanism of action of this compound as a non-competitive inhibitor.

[0271] Example 51

[0272] The pathogenesis of various neurodegenerative diseases, such as Alzheimer's and Parkinson's, is closely related to neuroinflammation. Microglia in the brain are overactivated under pathological signals, releasing large amounts of pro-inflammatory factors such as interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) through inflammatory signaling pathways such as NLRP3 / IL-1β. As a core inflammatory mediator, IL-1β can not only directly induce neuronal damage and apoptosis, but also further amplify the neuroinflammatory response, disrupt the integrity of the blood-brain barrier, and exacerbate core pathological processes of the disease (such as Aβ deposition and Tau protein phosphorylation). Therefore, inhibiting IL-1β-related inflammatory responses has become an important strategy for treating neurodegenerative diseases.

[0273] Microglia can be activated and highly express IL-1β upon stimulation with lipopolysaccharide (LPS). Therefore, this model is often used in vitro to simulate neuroinflammatory states and as an experimental system for screening small molecules with anti-neuroinflammatory drug activity.

[0274] In this embodiment, an LPS-induced mouse microglia (BV2) inflammation model was used to determine the effect of the 48 synthesized compounds on inhibiting the inflammatory response of BV2 cells.

[0275] The mouse microglia (BV2) cells used in the experiment were provided by Shanghai CyberKang Biotechnology Co., Ltd.

[0276] BV2 cells in the logarithmic growth phase were digested with trypsin, counted, and then the cell concentration was adjusted to 2 × 10⁻⁶ cells / cells. 4 / wells were seeded into 96-well plates and cultured overnight at 37°C in a 5% CO2 incubator to allow the cells to adhere.

[0277] The experiment was divided into a blank control (Control) group, a model control (LPS) group, and a sample group. The Control group and LPS group were given 100 μL / well of complete culture medium containing an equal volume of DMSO, while the sample group was given 100 μL / well of working solution containing different concentrations of the test compound.

[0278] After continuous culture for 2 h, 11 μL / well of blank complete medium was added to the Control group, and 11 μL / well of LPS solution with a concentration of 10 μg / mL was added to the LPS group and the sample group, followed by continuous culture for 24 h. Each treatment group was replicated 3 times.

[0279] After the culture ended, the cell supernatant was collected and the concentration of IL-1β in the supernatant was measured according to the operating instructions of the IL-1β ELISA detection kit. The results are shown in Table 4.

[0280]

[0281] In the table a-e represents the compound concentration: a 10 μM, b 5 μM, c 2.5 μM, d 2 μM, e 1 μM; f IC 50 (μM): The compound concentration corresponding to when IL-1β is inhibited by half.

[0282] As can be seen from the results in Table 4, at the tested concentrations, 41 compounds had an inhibition rate higher than 50% on LPS-induced IL-1β production, and higher than donepezil (44.15 ± 1.34%), showing good anti-neuroinflammatory activity. Among them, the compound JOPQ in Example 14 had an IC 50 value of 0.17 ± 0.02 μM for inhibiting IL-1β production.

[0283] Example 52

[0284] The male ICR mice (6 - 8 weeks old, weighing 20 - 25 g) used in this experiment were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Production License No. SCXK (Beijing) 2021 - 0006). All animal experiment operations were carried out in accordance with relevant national regulations and approved by the Animal Ethics Committee.

[0285] After 56 mice were adaptively fed under standard conditions for 1 week, they were randomly divided into 7 groups of​​Except for the normal control group and the solvent control group, all other groups were equipped with an Alzheimer's disease (AD) mouse model by intraperitoneal injection of scopolamine (SCOP, 4 mg / kg), which was administered 30 minutes before the behavioral test. The solvent control group was injected with an equal volume of physiological saline, and the normal group was not treated in any way.

[0287] Each group was given the corresponding drug or solvent by gavage 30 minutes before modeling. The normal control group and the solvent control group were given the same volume of solvent by gavage.

[0288] The novel object recognition experiment was used to evaluate the effects of the JOPQ on environmental perception and short-term memory in mice. The experimental setup was a 50cm × 50cm × 50cm open box. On Day 1, mice were placed alone in an empty box and allowed to explore freely for 5 minutes to acclimatize. On Day 2, two identical objects (A1 and A2) were placed in the box, and the mice were placed facing the box wall. The exploration time for each object was recorded for 5 minutes. On Day 3, one of the objects was replaced with a new object (B), and the exploration time for both the familiar object (A) and the new object (B) was recorded for 5 minutes. After each experiment, the box and objects were wiped with 75% ethanol to eliminate odor interference.

[0289] The Discrimination Index (DI) is calculated using the formula: DI = Time to explore new objects / (Time to explore familiar objects + Time to explore new objects) × 100%.

[0290] The experimental results are shown in Table 5 and Figure 2 As shown. Figure 2 The image shows a heatmap of a mouse's trajectory when exploring objects. The white square in the upper left corner represents a familiar object, and the red circle in the lower right corner represents a new object. As the time spent in the object increases, the mouse's trajectory changes from light green to dark red.

[0291]

[0292] Compared with the normal control group, the model group mice showed a significant decline in environmental perception and short-term memory, manifested as a significant decrease in the discrimination index of new objects. P <0.01), indicating that scopolamine induces short-term memory impairment.

[0293] Compared with the model group, mice in the low, medium, and high dose groups of JOPQ showed significantly improved discrimination indices for novel objects. P The results showed that JOPQ (<0.01) and exhibited a certain dose-dependent effect, indicating that JOPQ can significantly improve the model mice's ability to perceive the environment and their short-term memory.

[0294] The Morris water maze test was used to evaluate the effects of the JOPQ on spatial learning and long-term memory in mice. The apparatus consisted of a circular pool with a diameter of 130 cm and a height of 60 cm, with the water temperature maintained at 24 ± 1 °C. Spatial cues were marked on the pool walls. A hidden platform with a diameter of 8 cm was placed in the pool, 1 cm below the water surface. A camera system automatically recorded the time it took for the mice to reach the platform (escape latency) and the number of times they crossed the platform while swimming.

[0295] Memory training was conducted on days 1-6 of the experiment. The mice were given the drug 1 hour before training and the model was established 30 minutes before training. During training, the mice in each group entered the water with their abdomen facing the opposite quadrant of the platform. Each training session lasted 90 seconds. If the mice did not reach the platform within 90 seconds, they were guided to stay on the platform for 15 seconds.

[0296] On day 7, the underwater platform was removed, and all other procedures were the same. The escape latency of the mice and the number of times they crossed the virtual platform (the original underwater platform location) within 90 seconds were recorded. The results are as follows: Figure 3 As shown in Table 6.

[0297]

[0298] The Morris water maze test results showed that, compared with the normal group, the model group mice had impaired spatial learning and long-term memory abilities, manifested as a significant increase in escape latency. P <0.01), the number of times entering the virtual platform decreased significantly ( P <0.01), and the degree of disorder in movement trajectories increased significantly. Compared with the model group, the escape latency of mice in the low- and medium-dose JOPQ groups was significantly reduced ( P <0.05), the escape latency was significantly reduced in the high-dose group of mice ( P <0.01), and in addition, compared with the model group, the number of times mice in the high-dose JOPQ group entered the virtual platform was significantly increased ( P <0.01), and superior to donepezil ( P <0.05). The above results indicate that JOPQ can significantly improve the learning and long-term memory abilities of scopolamine model mice.

[0299] After the behavioral experiment, mouse brain tissue was collected. The acetylcholine content in mouse hippocampus was detected using a mouse acetylcholine content detection kit, and the IL-1β, IL-6 and TNFα content in mouse hippocampus was detected using an ELISA kit.

[0300]

[0301] Table 7 shows that, compared with the normal group, the ACh level in the hippocampus of the model group mice was significantly decreased ( P<0.01), mice exhibited impaired learning and memory abilities. Compared to the model group, ACh levels were significantly increased in the medium and high dose JOPQ groups ( P <0.01).

[0302] Furthermore, compared with the normal group, the levels of IL-1β, IL-6, and TNFα in the hippocampus of mice in the model group were significantly increased. P <0.01, P <0.01, P <0.01). Compared with the model group, the IL-6 levels in the low, medium, and high dose groups of JOPQ were significantly reduced ( P <0.01, P <0.01, P <0.01), and the JOPQ medium and high dose groups were superior to donepezil ( P <0.01, P <0.01). Compared with the model group, the IL-1β levels in the JOPQ medium and high dose groups were significantly reduced ( P <0.05, P <0.01), while there was no statistically significant difference between the donepezil group and the model group. Compared with the model group, the high-dose JOPQ group significantly reduced TNFα levels ( P <0.01), and superior to donepezil ( P <0.01).

[0303] The embodiments described above are merely specific examples of the present invention and are not intended to limit the scope of protection of the present invention. Various changes, modifications, or substitutions made to these embodiments by those skilled in the art without departing from the principles and spirit of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A class of compounds having anti-neuroinflammatory activity and selective inhibition of butyrylcholinesterase activity, which are compounds having the structure shown in the following formula (I) and pharmaceutically acceptable salts thereof: ； wherein: R1 is hydrogen or C1-C4 alkyl; R2 is substituted amino or 5, 6-membered nitrogen heterocyclic group.

2. The compound according to claim 1, wherein R1 is hydrogen, methyl, ethyl, propyl, n-butyl or t-butyl; and R2 is N,N-dimethylamino, N-ethyl-N-methylamino, N,N-diethylamino, N,N-diisopropylamino, N,N-di(2-chloroethyl)amino, pyrrolidin-1-yl, piperidin-1-yl or morpholin-4-yl. R1 is hydrogen, methyl, ethyl, propyl, n-butyl or t-butyl; and R2 is N,N-dimethylamino, N-ethyl-N-methylamino, N,N-diisopropylamino, N,N-di(2-chloroethyl)amino, pyrrolidin-1-yl or piperidin-1-yl. R1 is hydrogen, methyl, ethyl, n-butyl or t-butyl; and R2 is N,N-dimethylamino, N-ethyl-N-methylamino, N,N-diisopropylamino, pyrrolidin-1-yl or piperidin-1-yl.

3. The compound of claim 1, wherein R1 is hydrogen, methyl, n-butyl or t-butyl; and R2 is N,N-diisopropylamino, pyrrolidin-1-yl or piperidin-1-yl.

4. The compound of claim 1, wherein R1 is methyl or n-butyl, and R2 is piperidin-1-yl.

5. The compound of claim 1, wherein 7. Use of a compound according to any one of claims 1 to 6 for the manufacture of a medicament for the prevention and / or treatment of neurodegenerative diseases.

6. The compound of claim 1, wherein 8. The use according to claim 7, wherein the neurodegenerative disease comprises Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis or spinocerebellar ataxia.

9. The use according to claim 7, wherein the neurodegenerative disease is selected from Alzheimer's disease or Parkinson's disease.

10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof as an active ingredient, together with one or more pharmaceutically acceptable carriers or excipients. ​ ​

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