Benzimidazole-amino furazan-oxadiazole skeleton compound as well as preparation method and application thereof

Abstract

The invention discloses a compound with a benzimidazole-amino furazan-oxadiazole skeleton as well as a preparation method and application of the compound. The structure of the compound is shown in general formulas I and II. According to the present invention, butyrylcholine esterase inhibition activity, Nrf2 activation activity, in vitro pharmacodynamic experiment and Morris water maze experiment are adopted as carriers to evaluate the treatment of Alzheimer's disease (especially moderate and severe Alzheimer's disease) by using the compounds represented by the general formulas I and II, and the results show that the compounds have good in vitro and in vivo activity and extremely high selectivity; the compound can be used as a candidate lead compound for further development of an anti-Alzheimer's disease effect by selectively inhibiting butyrylcholine esterase and activating Nrf2.

Description

Technical Field

[0001] This invention relates to compounds, their preparation methods and uses, particularly benzimidazole-aminofuran-oxadiazole skeletal compounds and their preparation methods and uses. Background Technology

[0002] The Disease Burden and Treatment Challenges of Alzheimer's Disease (AD). Alzheimer's disease (AD) is a neurodegenerative disease characterized by progressive cognitive decline. Its core pathological features include β-amyloid (Aβ) plaque deposition in the brain, neurofibrillary tangles formed by highly phosphorylated Tau protein, synaptic dysfunction, and depletion of neurotransmitters, including acetylcholine (ACh). The "2023 China Alzheimer's Disease Data and Prevention Strategies" report shows that the prevalence of AD in my country among people aged 60 and above is 3.94%, with approximately 11.83 million patients, accounting for about a quarter of the global total. Furthermore, AD is the fifth leading cause of death in China. Predictive models warn that the number of AD patients and the accompanying enormous direct economic burden will continue to rise in the coming decades. Currently approved therapies (including cholinesterase inhibitors donepezil, rivastigmine, galantamine, and huperzine A, NMDA receptor antagonist memantine, and gut-brain axis modulator mannostatin, etc.) can only temporarily and to a limited extent relieve some symptoms and cannot reverse the disease progression; their overall efficacy is far from clinically expected. While the recently approved Aβ-targeting antibodies aducanumab and lecanemab in the US are considered breakthroughs in mechanism, their high cost and significant safety risks, such as amyloid-related imaging abnormalities (ARIA), have sparked widespread controversy and highlighted the significant unmet needs in the treatment of Alzheimer's disease (AD).

[0003] Advantages and Necessity of Multi-Target Therapy Strategies (MTDLs). Alzheimer's disease (AD) is pathologically complex, involving multiple intertwined factors such as amyloid cascade, aberrant Tau phosphorylation, neuroinflammation, oxidative stress, cholinergic system failure, metal ion metabolism disorders, mitochondrial dysfunction, and ferroptosis. Previous single-mechanism drug development strategies, primarily based on a "one-drug, one-target" approach, have repeatedly failed, indicating that targeting a single pathological link is insufficient for effectively intervening in this systemically complex disease. In contrast, the multi-target directed ligand (MTDL) design strategy, by simultaneously modulating multiple key pathogenic pathways through a single molecular entity, can achieve additive or even synergistic therapeutic effects. Compared to targeted specific drug combinations (TSDCs), MTDLs offer significant advantages such as no risk of drug interactions, a single pharmacokinetic profile (avoiding pharmacokinetic differences between components), simplified dosing regimens (improving patient compliance), and a reduction in potential adverse reactions. The core challenges in developing MTDLs lie in the rational selection of target combinations (ensuring regulation of related and concurrent pathological processes), optimization of molecular design (connecting, fusing, or merging different pharmacophores), and drugability considerations (such as ensuring blood-brain barrier permeability: requiring molecules to have a suitable balance between lipid solubility and water solubility). Especially for AD, MTDL molecules need to achieve a balanced activity intensity among various targets to achieve optimal synergistic pharmacological effects.

[0004] BChE and Nrf2: Dual Targets and Their Synergistic Mechanism. Traditional cholinergic therapy relies on inhibiting acetylcholinesterase (AChE). However, in the later stages of Alzheimer's disease (AD), neuronal loss weakens AChE's effect, resulting in a clinical efficacy period of less than 12 months for AChE inhibitors (AChEIs), and long-term use is prone to causing peripheral cholinergic side effects. Butyrylcholinesterase (BChE) activity increases compensatorily in the later stages of AD, becoming the main enzyme for hydrolyzing acetylcholine. Therefore, inhibiting BChE (BChEIs) not only increases acetylcholine levels in the brain and improves cognition, but also has the advantages of longer duration of action and lower risk of peripheral side effects compared to AChEIs. Studies have also shown that BChE participates in regulating multiple pathological processes, including combating Aβ deposition, alleviating Tau pathology, and relieving oxidative stress and neuroinflammation, and is considered a target with multiple therapeutic potentials. Nrf2 is a core transcription factor regulating cellular antioxidant stress and anti-inflammatory responses. Activation of Nrf2 can induce the expression of multiple protective genes, effectively reduce oxidative damage and inflammation, and play an important role in maintaining mitochondrial function and metal ion homeostasis. Numerous studies have confirmed the therapeutic potential of the Nrf2 pathway in neurodegenerative disease models.

[0005] Structure-activity relationship studies have successfully identified a series of novel BChE inhibitors with high inhibitory activity and excellent selectivity, including benzimidazole-oxadiazole derivatives. Building upon this solid foundation, this invention focuses on and utilizes benzimidazole-aminofuran as the core structural framework for the design and development of dual-target (BChE-Nrf2) regulators. The advantage of this structure lies in the fact that previous derivatives have demonstrated that the benzimidazole-aminofuran structure can effectively and selectively inhibit BChE, possessing the potential to become a potent BChE pharmacophore. The benzimidazole and aminofuran ring system has multiple modifiable sites, facilitating the rational introduction of pharmacophores regulating the Nrf2 pathway, making it an ideal platform for achieving "dual-target integrated" MTDL design. Summary of the Invention

[0006] Objective of the Invention: This invention provides an innovative treatment strategy for Alzheimer's disease by designing and developing novel compounds based on the benzimidazole-aminofuran structure. These compounds achieve dual synergistic pharmacological effects by simultaneously and efficiently inhibiting butyrylcholinesterase (BChE) and activating the nuclear factor E2-related factor 2 (Nrf2) signaling pathway: (1) increasing intracranial acetylcholine levels, providing sustained improvement in cognitive function while avoiding peripheral cholinergic side effects; (2) significantly enhancing the anti-ferroptosis ability of nerve cells and providing neuroprotection by regulating the ferroptosis defense network through the Nrf2-GSH-GPX4 pathway. This structural framework has been demonstrated to possess excellent BChE inhibitory activity and selectivity, providing a powerful technical solution to overcome the limitations of current single-target treatments for AD. Another objective of this invention is to provide a method for preparing the above-mentioned compounds and their uses.

[0007] Technical solution: The compounds of the benzimidazole-aminofuran-oxadiazole skeleton or their pharmaceutically acceptable salts as described in this invention have the following structures:

[0008]

[0009] in,

[0010] R 1 R 2 Each independently represents hydrogen or R 3 Represents hydrogen, methyl, or halogen; X represents carbon or nitrogen; n = 1, 2, 3; R 4 Represents hydrogen, C 1～6 Alkyl, substituted or unsubstituted phenyl, phenylalkyl; the substitution is selected from halogen, nitro, methoxy, C 1～6 Alkyl, halomethyl, cyclohexyl; the alkyl group in the phenylalkyl group is C10. 1～6 Alkyl group. * indicates a junction.

[0011] In this invention, phenylalkyl refers to an alkyl group directly attached to the oxadiazole ring, such as the structure of compound 43; substituted phenyl refers to a phenyl group attached to the oxadiazole ring, such as the structure of compound 20.

[0012] The aforementioned compounds with the benzimidazole-aminofuran-oxadiazole skeleton or pharmaceutically acceptable salts thereof,

[0013] R 1 R 2 Each independently represents hydrogen or R 3 Represents hydrogen, methyl, or bromine; X represents carbon or nitrogen; n = 1, 2; R 4 Represents hydrogen, C 1～6 Alkyl, substituted or unsubstituted phenyl, benzoalkyl; the substitution is selected from fluorine, chlorine, bromine, iodine, nitro, methoxy, C 1～6 Alkyl, trifluoromethyl, cyclohexyl; the alkyl group in the phenylalkyl group is C10. 1～6 alkyl.

[0014] The compound with the benzimidazole-aminofuran-oxadiazole skeleton or a pharmaceutically acceptable salt thereof is any one of the following compounds or a pharmaceutically acceptable salt thereof:

[0015]

[0016]

[0017]

[0018]

[0019]

[0020] The compound of the benzimidazole-aminofuran-oxadiazole skeleton or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt is a salt formed by the compound and any one of the following acids: hydrochloric acid, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, malic acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, citric acid, benzenesulfonic acid, p-toluenesulfonic acid, or ferulic acid.

[0021] The preparation method of the compound with the benzimidazole-aminofuran-oxadiazole skeleton or its pharmaceutically acceptable salt, wherein the preparation method of the compound represented by general formula I includes the following steps:

[0022]

[0023] The method for preparing the compound with the benzimidazole-aminofuran-oxadiazole skeleton or a pharmaceutically acceptable salt thereof, to... Using the raw material as a starting material, it was dissolved in acetic acid / water solution under ice bath conditions. Sodium nitrite was added, and the mixture was stirred at room temperature to obtain an intermediate. The intermediate is then refluxed with hydroxylamine hydrochloride and potassium hydroxide in diethylene glycol dimethyl ether at 100-120°C to obtain the intermediate. by Using chloroacetyl chloride or chloropropionyl chloride and sodium bicarbonate as starting materials, the mixture is stirred in dichloromethane at room temperature to obtain an intermediate. Then with intermediates Cesium carbonate was refluxed in N,N'-dimethylformamide at 40-60°C to give an intermediate. The intermediate was then refluxed with hydroxylamine hydrochloride and potassium carbonate in anhydrous ethanol at 70-90°C to obtain the intermediate. Finally and The target compound was obtained by amide condensation.

[0024] The above R 3 X,n is defined as described above.

[0025] The preparation method of the compound with the benzimidazole-aminofuran-oxadiazole skeleton or its pharmaceutically acceptable salt, wherein the preparation method of the compound represented by general formula II includes the following steps:

[0026]

[0027] The method for preparing the compound with the benzimidazole-aminofuran-oxadiazole skeleton or a pharmaceutically acceptable salt thereof, to... Using the raw material as a starting material, it was dissolved in acetic acid / water solution under ice bath conditions. Sodium nitrite was added, and the mixture was stirred at room temperature to obtain an intermediate. The intermediate is then refluxed with hydroxylamine hydrochloride and potassium hydroxide in diethylene glycol dimethyl ether at 100-120°C to obtain the intermediate. by Using chloroacetyl chloride and sodium bicarbonate as starting materials, the mixture was stirred in dichloromethane at room temperature to obtain an intermediate. intermediate The intermediate was obtained by reflux of cesium carbonate in N,N'-dimethylformamide at 40-60°C. The intermediate was then refluxed with hydroxylamine hydrochloride and potassium carbonate in anhydrous ethanol at 70-90°C to obtain the intermediate. Finally with R 4 The target compound was obtained by amide condensation of -COOH.

[0028] The above R 4 Defined as described above.

[0029] Pharmaceutical compositions comprising compounds of the benzimidazole-aminofuran-oxadiazole skeleton or pharmaceutically acceptable salts thereof and pharmaceutically acceptable carriers thereof.

[0030] Use of the compound of the benzimidazole-aminofuran-oxadiazole skeleton or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of Alzheimer's disease.

[0031] Furthermore, the pharmaceutical composition is formulated as tablets, capsules, powders, syrups, liquids, suspensions, or injections.

[0032] Beneficial Effects: This invention provides novel benzimidazole-aminofuran-oxadiazole derivatives represented by formulas I and II. Evaluation of the following key technical indicators confirmed the therapeutic potential of these compounds in treating Alzheimer's disease (AD), particularly moderate to severe AD: in vitro tests showed that the compounds exhibit highly efficient and selective inhibition of BChE; in vitro and cell model experiments demonstrated that the compounds effectively activate the Nrf2 signaling pathway; and in vivo behavioral evaluation methods, such as the Morris water maze test, confirmed that administration of the compounds significantly improved cognitive dysfunction in AD model animals. In summary, the compounds of this invention possess multi-target regulatory properties. This dual mechanism of action holds promise for simultaneously intervening in multiple pathological stages of AD (such as cholinergic system damage, oxidative stress, and neuroinflammation), thereby achieving the therapeutic goals of alleviating brain damage and delaying disease progression. Furthermore, the benzimidazole-aminofuran-oxadiazole heterocyclic skeleton helps maintain a reasonable molecular weight, suitable lipid-water partition coefficient, and favorable polar surface area. Based on the simulation predictions of the above physicochemical properties, the compound exhibits excellent blood-brain barrier penetration. Therefore, the compound of the present invention is a promising candidate lead compound for combating Alzheimer's disease (AD) through a dual mechanism (selective inhibition of BChE and activation of Nrf2). Attached Figure Description

[0033] Figure 1 This is a graph showing the results of a luciferase assay.

[0034] Figure 2 These are the results of oxidative stress ROS detection under a fluorescence microscope;

[0035] Figure 3 This is the result of a mouse behavioral study. Detailed Implementation

[0036] Example 1

[0037] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-bromo-5-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0038] (1) Synthesis of (E)-N-hydroxy-1H-benzo[d]imidazolium-2-formamidinium (intermediate 1-1)

[0039] Under ice bath conditions, 10 g of 2-(1H-benzo[d]imidazol-2-yl)acetonitrile was dissolved in 50 mL of acetic acid, and 40 mL of water was added. 5 g of sodium nitrite was dissolved in 10 mL of water and poured into a constant pressure dropping funnel. This solution was then slowly added dropwise to the acetic acid / water solution of 2-(1H-benzo[d]imidazol-2-yl)acetonitrile, and the mixture was stirred at room temperature for 0.5 h. A saturated sodium bicarbonate solution was added to the reaction mixture to adjust the pH to alkaline, resulting in the precipitation of a brownish-yellow solid. The solid was filtered, washed three times with water, and then three times with isopropyl ether. After drying, a brownish-yellow solid was obtained.

[0040] (2) Synthesis of 4-(1H-benzo[d]imidazol-2-yl)-1,2,5-oxadiazol-3-amine (intermediates 1-2)

[0041] Dissolve 15.4 g of potassium hydroxide in 100 mL of water and cool to room temperature. Dissolve 13.2 g of hydroxylamine hydrochloride in 100 mL of water. While stirring, add the potassium hydroxide solution dropwise to the hydroxylamine hydrochloride solution. Add (E)-N-hydroxy-1H-benzo[d]imidazolium-2-formamidinium (intermediate 1) and diethylene glycol dimethyl ether (200 mL). Reflux at 115 °C for 6 h. The solution changes from clear to having a solid precipitate. After the reaction is complete, cool to room temperature. Filter and wash three times with isopropyl ether. Dry to obtain a brownish-yellow solid.

[0042] (3) Synthesis of N-(3-bromo-5-cyanophenyl)-2-chloroacetamide (intermediate 1-3a)

[0043] 3-Amino-5-bromobenzonitrile and sodium bicarbonate were dissolved in dichloromethane, and chloroacetyl chloride was slowly added dropwise while stirring at room temperature for 1 hour. After the reaction was complete, dichloromethane was added to dilute the reaction solution, and the mixture was washed three times with saturated sodium bicarbonate solution. The dichloromethane layer was collected. The product in the saturated sodium bicarbonate layer was then extracted with dichloromethane, and the two dichloromethane layers were combined. The solvent was removed by evaporation under reduced pressure to obtain a milky white solid, which could be directly added to the next step without further purification.

[0044] (4) Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-bromo-5-cyanophenyl)acetamide (intermediate 1-4a)

[0045] 4-(1H-benzo[d]imidazol-2-yl)-1,2,5-oxadiazol-3-amine (intermediate 2) was dissolved in N,N'-dimethylformamide, and N-(3-bromo-5-cyanophenyl)-2-chloroacetamide (intermediate 3) and cesium carbonate were added. The mixture was refluxed at 50°C for 4 h. After the reaction was completed, the reaction solution was added dropwise to water while stirring, and a solid precipitated. The solid was filtered and dried to obtain a milky white solid, which could be directly added to the next step without purification.

[0046] (5) Synthesis of (Z)-2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-bromo-5-(N'-hydroxycarbonylamineimino)phenyl)acetamide (intermediate 1-5a)

[0047] 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-bromo-5-cyanophenyl)acetamide (intermediate 4), hydroxylamine hydrochloride, and potassium carbonate 3- were dissolved in anhydrous ethanol and refluxed at 85°C for 12 h. The solution changed from a milky white suspension to a pale yellow, clear, and transparent solution. The solution was filtered while hot, and pale yellow crystals precipitated upon standing. The crystals were then filtered again, washed with anhydrous ethanol, and dried to obtain a pale yellow solid, which was directly added to the next step without further purification.

[0048] (6) Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-bromo-5-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0049] 3,4-Difluorobenzoic acid, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, and N,N-diisopropylethylamine were dissolved in N,N'-dimethylformamide and stirred at room temperature for 30 min. After the activation of 3,4-difluorobenzoic acid was monitored by TLC, (Z)-2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-bromo-5-(N'-hydroxycarbonylamineimino)phenyl)acetamide (intermediate 5) was added to the reaction solution and stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was added dropwise to water while stirring, and a precipitate was formed. The precipitate was filtered and dried. The filter cake was dissolved in dimethyl sulfoxide, potassium hydroxide was added, and the mixture was stirred at room temperature for 1 h. After the reaction was completed by TLC, the reaction solution was added dropwise to water while stirring, and a precipitate was formed. The precipitate was filtered and dried. The filter cake was purified by thin-layer chromatography (developing solvent: petroleum ether: ethyl acetate: triethylamine = 60:60:1) to give a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-bromo-5-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO) δ11.18(d,J=18.0Hz,1H),8.43–7.65(m,8H),7.55–7.32(m,2H),7.04(s,2H),5.66(d,J=6.0Hz,2H).

[0050] Example 2

[0051] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0052] Referring to the synthesis method of Example 1, intermediates 1-3a in Example 1 were replaced with 2-chloro-N-(3-cyanophenyl)acetamide to obtain a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1H NMR(300MHz,DMSO-d6)δ10.89(s,1H),8.46(d,J=1.7Hz,1H),8.30–8.17(m,1H),8.15–7.99(m ,1H),7.94–7.65(m,5H),7.57(t,J=7.9Hz,1H),7.50–7.37(m,2H),7.03(s,2H),5.64(s,2H).

[0053] Example 3

[0054] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(4-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)pyridin-2-yl)acetamide

[0055] Referring to the synthesis method of Example 1, intermediates 1-3a in Example 1 were replaced with 2-chloro-N-(4-cyanopyridin-2-yl)acetamide, yielding a pale yellow solid, which is the synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(4-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)pyridin-2-yl)acetamide. 1 HNMR(300MHz,DMSO)δ11.63(s,1H),8.66(d,J=4.6Hz,2H),8.22(s,1H),7.93–7 .80(m,3H),7.61(d,J=4.6Hz,1H),7.52–7.34(m,3H),7.03(s,2H),5.72(s,2H).

[0056] Example 4

[0057] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(4-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0058] Referring to the synthesis method of Example 1, intermediates 1-3a in Example 1 were replaced with 2-chloro-N-(4-cyanophenyl)acetamide to obtain a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(4-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1H NMR (400MHz, DMSO-d6) δ10.91 (d, J=9.6Hz, 1H), 8.29–8.20 (m, 1H), 8.11–8. 00(m,3H),7.89–7.79(m,5H),7.50–7.36(m,2H),7.02(s,2H),5.64(s,2H).

[0059] Example 5

[0060] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)propionamide

[0061] Referring to the synthesis method of Example 1, intermediates 1-3a in Example 1 were replaced with 2-chloro-N-(3-cyanophenyl)propionamide to obtain a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)propionamide. 1 H NMR (400MHz, DMSO) δ10.26(s,1H),8.32–8.16(m,2H),8.07(d,J=4.5Hz,1H),7.91–7.63(m,5H),7.49(t,J=7 .9Hz,1H),7.42(t,J=7.3Hz,1H),7.33(t,J=7.5Hz,1H),7.02(s,2H),5.08–4.91(m,2H),3.07–2.91(m,2H). 13 CNMR(151MHz,DMSO)δ169.31,168.75,156.63,142.07,141.05,140.01,138.69,135.59,130.22,126.72,126.37,126.35,124.95,1 23.59,122.63,122.39,120.34,119.71,119.59,118.05,118.01,117.88,111.95,42.00,36.93.HRMS(ESI):found527.1392,calcd for C 25 H 18 N8O3[MH] - 527.1397

[0062] Example 6

[0063] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)-5-methylphenyl)acetamide

[0064] Referring to the synthesis method of Example 1, intermediates 1-3a in Example 1 were replaced with 2-chloro-N-(3-cyano-5-methylphenyl)acetamide, yielding a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)-5-methylphenyl)acetamide. 1 H NMR (400MHz, DMSO-d6) δ10.82(s,1H),8.29–8.16(m,2H),8.10–7.98(m,1H),7.94–7.82(m,2H),7. 79–7.65(m,1H),7.62(d,J=6.9Hz,2H),7.52–7.34(m,2H),7.03(s,2H),5.63(s,2H),2.38(s,3H). 13 C NMR (151MHz, DMSO) δ168.76,165.89,156.49,141.98,141.83,140.02,139.82,138.74,136.56,126.80,126.36,125.19,123.70,1 23.18,123.04,121.23,120.42,119.65,119.52,118.01,117.88,115.35,111.60,48.67,21.51.HRMS(ESI):found527.1387,calcd for C 25 H 18 N8O3[MH] - 527.1397

[0065] Example 7

[0066] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-phenyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0067] Referring to the synthesis method of Example 2, 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with benzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-phenyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO-d6)δ8.47(t,J=1.6Hz,1H),8.21–8.15(m,2H),7.92–7.70(m,5H),7.6 9–7.61(m,2H),7.56(t,J=7.9Hz,1H),7.51–7.36(m,2H),7.03(s,2H),5.68–5.59(m,2H). 13 C NMR (151MHz, DMSO) δ175.93,168.62,165.98,156.50,141.99,141.85,140.06,138.76,136.57,133.83,130.43,13 0.04,128.39,127.19,125.20,123.80,123.70,122.53,122.48,120.42,118.03,111.64,48.71.HRMS(ESI):found 477.1427,calcd for C 25 H 18 N8O3[MH] - 477.1429

[0068] Example 8

[0069] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(2-chlorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0070] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2-chlorobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(2-chlorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ8.44(s,1H),8.19–8.15(m,1H),7.98–7.68(m,6H),7.65–7.51(m,2H),7.48–7.32(m,2H),7.03(s,2H),5.69–5.58(m,2H).13 C NMR (151MHz, DMSO) δ174.51,168.37,166.00,156.49,141.98,141.83,140.06,138.75,136.56,134.74,132.80,132.65,131.79, 130.50,128.47,126.98,125.19,123.70,123.14,122.60,120.42,118.02,111.64,48.70.HRMS(ESI):found511.S27-1036,calcd for C 25 H 17 ClN8O3[MH] - 511.1039

[0071] Example 9

[0072] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(3-chlorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0073] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 3-chlorobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3-chlorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ10.90(s,1H),8.47(s,1H),8.16(d,J=11.7Hz,2H),7.85(m,J=19.2,9.0H z,5H),7.70(m,J=7.9Hz,1H),7.58(m,J=7.7Hz,1H),7.52–7.36(m,2H),7.04(s,2H),5.66(s,2H). 13C NMR (151MHz, DMSO) δ174.74,168.68,165.98,156.50,141.98,141.82,139.93,138.75,136.57,134.63,133.61,132.07,13 0.50,127.86,127.13,127.01,125.70,125.21,123.71,122.64,122.52,120.43,117.96,111.63,48.69.HRMS(ESI):found 511.1041,calcd for C 25 H 17 ClN8O3[MH] - 511.1039

[0074] Example 10

[0075] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(4-chlorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0076] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 4-chlorobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(4-chlorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO-d6)δ10.99(s,1H),8.49(s,1H),8.19(d,J=8.6Hz,2H),7.96– 7.68(m,6H),7.57(t,J=7.9Hz,1H),7.50–7.39(m,2H),7.03(s,2H),5.67(s,2H). 13 C NMR (151MHz, DMSO) δ175.09,168.66,165.97,156.49,141.98,141.83,139.91,138.74,138.68,136.53,130.45,130.23,13 0.20,127.05,125.23,123.73,122.64,122.60,122.50,120.43,118.00,111.59,48.67.HRMS(ESI):found511.1039,calcd for C 25 H 17 ClN8O3[MH]- 511.1039

[0077] Example 11

[0078] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(2-fluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0079] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2-fluorobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(2-fluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ8.44(s,1H),8.21(t,J=7.2Hz,1H),8.07–7.69(m,6H),7.61–7.38(m,4H),7.03(s,2H),5.70(d,J=30.3Hz,2H). 13 C NMR (151MHz, DMSO) δ172.98,168.32,165.96,161.25,159.54,156.48,141. 97,141.80,139.87,138.74,136.52,136.19,136.13,131.32,130.51,127.0 1,125.98,125.96,125.25,123.75,122.67,122.52,120.44,118.01,117.80 ,117.66,112.22,112.14,111.56,48.64.HRMS(ESI):found497.1485,calcd for C 25 H 17 FN8O3[M+H] + 497.1480

[0080] Example 12

[0081] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(3-fluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0082] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 3-fluorobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3-fluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO-d6)δ10.98(s,1H),8.46(s,1H),8.03(d,J=7.4Hz,1H),7.97–7.78(m, 5H),7.74–7.67(m,1H),7.63–7.53(m,2H),7.51–7.36(m,2H),7.04(s,2H),5.66(s,2H). 13 C NMR (151MHz, DMSO) δ174.88,168.70,165.98,163.47,161.84,156.50,141.98,141.83,139.93,138.74,136.55,132.48,132.42,130.47,127.0 1,125.76,125.70,125.21,124.75,123.71,122.64,122.54,120.90,120.76,120.43,117.96,115.17,115.01,111.61,48.67.HRMS(ESI):found 497.1484,calcd for C 25 H 17 FN8O3[M+H] + 497.1480

[0083] Example 13

[0084] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(4-fluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0085] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 4-fluorobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(4-fluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1H NMR(300MHz,DMSO-d6)δ10.79(s,1H),8.39–8.17(m,2H),7.98(s,1H),7.93–7. 82(m,2H),7.79(d,J=6.9Hz,1H),7.58–7.29(m,6H),7.03(s,4H),5.63(s,2H). 13 C NMR (151MHz, DMSO) δ175.10,168.61,165.97,164.58,156.50,141.98,141.83,139.90,138.75,136.57,131.36,131.30,13 0.47,127.14,125.21,123.72,122.60,122.45,120.54,120.43,117.99,117.41,117.26,111.63,48.68.HRMS(ESI):found 497.1485,calcd for C 25 H 17 FN8O3[M+H] + 497.1480

[0086] Example 14

[0087] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(2-nitrophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0088] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2-nitrobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(2-nitrophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO-d6)δ10.93(s,1H),8.41(s,1H),8.32–8.25(m,1H),8.22–8.16(m,1H),8.08– 7.97(m,1H),7.92–7.77(m,2H),7.64–7.55(m,4H),7.52–7.38(m,1H),7.03(s,2H),5.66(s,2H). 13C NMR (151MHz, DMSO) δ173.11,168.59,166.00,156.49,141.97,141.81,139.97,138.74,136.55,134.72,134.40,132.1 2,130.61,126.62,125.41,125.21,123.71,122.72,122.64,120.42,117.99,117.96,111.62,48.68.HRMS(ESI):found 524.1427,calcd for C 25 H 17 N9O5[M+H] + 524.1425

[0089] Example 15

[0090] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3-nitrophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0091] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 3-nitrobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3-nitrophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(400MHz,DMSO-d6)δ10.92(s,1H),8.81(s,1H),8.60–8.51(m,2H),8.46(s,1H),7.97–7 .76(m,5H),7.56(t,J=7.9Hz,1H),7.50–7.37(m,2H),7.04(s,2H),5.63(d,J=10.6Hz,2H). 13 C NMR (151MHz, DMSO) δ174.24,168.77,165.99,156.50,148.68,141.98,141.83,139.96,138.75,136.56,134.38,131.94,130.5 4,128.02,126.84,125.21,123.71,122.95,122.62,122.58,120.43,118.00,111.62,48.69.HRMS(ESI):found524.1421,calcd for C 25 H17 N9O5[M+H] + 524.1425

[0092] Example 16

[0093] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(4-nitrophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0094] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 4-nitrobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(4-nitrophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO-d6)δ11.31(s,1H),8.63–8.31(m,5H),7.93–7.76(m,4H),7 .56(t,J=8.0Hz,1H),7.50–7.36(m,2H),7.01(s,2H),5.64(d,J=36.9Hz,2H). 13 C NMR (151MHz, DMSO) δ174.32,168.91,165.99,156.45,150.40,141.95,141.84,139.95,138.71,136.40,130.45,129.95,12 9.05,126.74,125.30,125.04,123.79,122.69,122.65,120.42,118.00,111.50,48.65.HRMS(ESI):found522.1281,calcd for C 25 H 17 N9O5[MH] - 522.1280

[0095] Example 17

[0096] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(2-methoxyphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0097] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2-methoxybenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(2-methoxyphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (400MHz, DMSO-d6) δ10.88(s,1H),8.42(s,1H),8.10–8.02(m,1H),7.88(t,J=8.8Hz,1H),7.84–7.76(m,2H),7.72–7.65(m,2H),7.55(t ,J=7.9Hz,1H),7.49–7.43(m,1H),7.43–7.37(m,1H),7.32(d,J=8.5Hz,1H),7.17(t,J=7.6Hz,2H),7.03(s,2H),5.64(s,2H),3.95(s,3H). 13 C NMR (151MHz, DMSO) δ175.56,167.88,165.94,158.65,156.50,141.98,141.82,139.87,138.74,136.57,135.30,131.73,130.4 2,127.39,125.20,123.70,122.59,122.29,121.35,120.42,117.95,113.42,112.74,111.63,56.63,48.69.HRMS(ESI):found 509.1681,calcd forC 26 H 20 N8O4[M+H] + 509.1680

[0098] Example 18

[0099] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(3-methoxyphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0100] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 3-methoxybenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3-methoxyphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1H NMR(300MHz,DMSO-d6)δ11.35(s,1H),8.49(s,1H),7.92–7.73(m,5H),7.67–7.52(m, 3H),7.50–7.37(m,2H),7.31(d,J=8.1Hz,1H),7.05(s,2H),5.71(s,2H),3.88(s,3H). 13 C NMR (151MHz, DMSO) δ175.79,168.65,165.98,160.17,156.49,141.98,141.89,140.01,138.74,136.47,131.37,130.34,127.1 0,125.22,124.92,123.72,122.62,122.52,120.75,120.41,120.04,117.92,112.82,111.60,56.03,48.71.HRMS(ESI):found 509.1680,calcd for C 26 H 20 N8O4[M+H] + 509.1680

[0101] Example 19

[0102] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(4-methoxyphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0103] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 4-methoxybenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(4-methoxyphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (400MHz, DMSO) δ10.99(s,1H),8.44(s,1H),8.16–8.02(m,2H),7.95–7.68(m,4H),7.59–7.37(m, 4H),7.17(d,J=8.9Hz,1H),7.12–6.99(m,2H),5.70–5.57(m,2H),3.86(d,J=7.5Hz,2H),2.55(s,3H). 13C NMR (151MHz, DMSO) δ175.77,168.43,165.94,165.80,163.59,156.50,141.98,141.84,139. 87,139.50,138.74,138.73,136.54,136.52,131.66,130.41,130.36,129.92,127.36,125. 21,125.19,123.71,123.69,122.57,122.34,121.95,121.79,120.42,117.97,116.79,116. 12,115.46,114.47,111.59,56.11,55.95,48.68,48.64.HRMS(ESI):found509.1684,calcd for C 26 H 20 N8O4[M+H] + 509.1680

[0104] Example 20

[0105] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(2-methylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0106] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2-methylbenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(2-methylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO) δ8.41(s,1H),8.10(d,J=6.8Hz,1H),7.96–7.72(m,4H),7.66–7.30(m,6H),7.01(s,2H),5.62(d,J=9.2Hz,2H),2.80–2.64(m,3H). 13C NMR (151MHz, DMSO) δ176.51,168.21,165.96,156.48,141.96,139.88,139.02,138.74,136.52,133.22,132.46,130.47,1 30.39,127.34,127.13,125.25,123.75,123.07,122.67,122.45,120.44,117.96,111.58,48.65,21.76.HRMS(ESI):found 493.1726,calcd for C 26 H 20 N8O3[M+H] + 493.1731

[0107] Example 21

[0108] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(3-methylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0109] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 3-methylbenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3-methylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO)δ11.09(d,J=53.7Hz,1H),8.49(s,1H),8.14–7.65(m,6H ),7.59–7.29(m,5H),7.03(d,J=4.1Hz,2H),5.72–5.60(m,2H),2.55(s,3H). 13 C NMR (151MHz, DMSO) δ176.04,168.57,165.97,156.49,141.97,141.81,139.89,139.66,138.74,136.55,134.51,130.45,129.9 5,128.64,127.23,125.55,125.23,123.73,123.69,122.60,122.42,120.43,117.98,111.60,48.67,21.22.HRMS(ESI):found 493.1577,calcd for C 26 H 20N8O3[MH] - 493.1586

[0110] Example 22

[0111] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(4-methylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0112] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 4-methylbenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(4-methylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO) δ10.99 (s, 1H), 8.48 (s, 1H), 8.10 (t, J = 13.5Hz, 2H), 7. 99–7.75(m,4H),7.63–7.36(m,5H),7.05(s,2H),5.67(s,2H),2.43(s,3H). 13 C NMR (151MHz, DMSO) δ175.99,168.52,165.95,156.50,147.61,144.29,141.98,141.84,139.90,138.74,136.55,130.58,130.4 0,128.35,127.27,125.21,123.71,122.57,122.38,121.06,120.42,117.98,111.61,107.20,48.68,21.71.HRMS(ESI):found 493.1729,calcd for C 26 H 20 N8O3[M+H] + 493.1731

[0113] Example 23

[0114] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(2-ethylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0115] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2-ethylbenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(2-ethylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO-d6)δ8.41(s,1H),8.06(d,J=6.8Hz,1H),7.99–7.77(m,4H),7.68–7 .32(m,6H),7.03(s,2H),5.74–5.57(m,2H),3.18–2.99(m,2H),1.22(t,J=7.5Hz,3H). 13 C NMR (151MHz, DMSO) δ176.37,168.31,165.97,156.49,145.07,141.97,141.84,139.95,138.74,136.56,133.40,131.02,130.67,1 30.47,127.31,127.15,125.20,123.71,122.61,122.51,122.48,120.42,117.96,111.63,48.69,27.29,16.02.HRMS(ESI):found 505.1741,calcd for C 27 H 22 N8O3[MH] - 505.1742

[0116] Example 24

[0117] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(4-ethylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0118] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 4-ethylbenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(4-ethylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1H NMR (300MHz, DMSO-d6) δ8.47(s,1H),8.14–7.73(m,6H),7.63–7.32(m,5H),7.06(s,2H),5.67(s,2H),2.70(s,2H),1.21(s,3H). 13 C NMR (151MHz, DMSO) δ175.95,168.52,165.94,156.51,150.23,141.99,141.84,139.92,138.75,136.55,130.37, 129.36,128.43,127.29,125.20,123.70,122.56,122.38,121.29,120.42,118.00,111.59,48.68,28.68,15.45.

[0119] Example 25

[0120] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(4-propylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0121] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 4-propylbenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(4-propylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO-d6)δ8.45(s,1H),8.08(d,J=8.2Hz,2H),7.94–7.67(m,4H),7.62–7.32(m,5H) ,7.02(s,2H),5.71–5.54(m,2H),2.67(t,J=7.5Hz,2H),1.70–1.53(m,2H),0.91(t,J=7.3Hz,3H). 13C NMR (151MHz, DMSO) δ176.01,168.52,165.96,156.49,148.74,141.98,141.82,139.90,138.74,136.55,130.44,130.00,128. 40,127.28,125.23,123.73,122.59,122.40,121.33,120.43,117.98,111.61,48.67,37.63,24.11,14.01.HRMS(ESI):found 521.2047,calcd forC 28 H 24 N8O3[M+H] + 521.2044

[0122] Example 26

[0123] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(2-isopropylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0124] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2-isopropylbenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(2-isopropylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ11.32(s,1H),8.45(s,1H),7.94(d,J=7.8Hz,1H),7.91–7.84(m,3H),7.80(d,J=7.6Hz,1H),7.71–7.61( m,2H),7.56(t,J=7.8Hz,1H),7.50–7.36(m,4H),7.04(s,2H),5.69(d,J=10.7Hz,2H),3.84–3.72(m,1H),1.27(d,J=6.7Hz,6H). 13C NMR (151MHz, DMSO) δ176.62,168.32,165.96,156.48,149.34,141.97,141.89,139.99,138.73,136.46,133.37,130.64,130. 38,127.20,126.91,125.22,123.72,122.63,122.53,122.40,120.41,117.94,111.59,48.70,29.87,24.04.HRMS(ESI):found 521.2046,calcd for C 28 H 24 N8O3[M+H] + 521.2044

[0125] Example 27

[0126] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(3-isopropylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0127] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 3-isopropylbenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3-isopropylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO-d6)δ10.92(s,1H),8.45(s,1H),8.01(d,J=11.0Hz,2H),7.95–7.77(m,4H),7.68– 7.52(m,3H),7.51–7.36(m,2H),7.05(s,2H),5.66(s,2H),3.19–2.91(m,1H),1.27(d,J=6.8Hz,6H). 13C NMR (151MHz, DMSO) δ176.10,168.57,165.96,156.50,150.38,141.98,141.82,139.90,138.75,136.57,131.99,130.45,130.14,127.25, 126.12,126.05,125.21,123.81,123.71,122.63,122.41,120.43,117.94,111.63,48.69,33.72,24.09.HRMS(ESI):found521.20,calcd for C 28 H 24 N8O3[M+H] + 521.2044

[0128] Example 28

[0129] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(4-isopropylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0130] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 4-isopropylbenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(4-isopropylphenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (500MHz, DMSO-d6) δ10.87(s,1H),8.45(s,1H),8.12(d,J=8.0Hz,2H),7.94–7.85(m,2H),7.85–7.77(m,2H),7.61–7.5 2(m,3H),7.48(t,J=7.5Hz,1H),7.42(t,J=7.3Hz,1H),7.03(s,2H),5.65(s,2H),3.10–2.95(m,1H),1.27(d,J=6.8Hz,6H). 13C NMR (126MHz, DMSO) δ175.99,168.56,165.99,156.52,154.80,142.00,141.85,139.92,138.77,138.69,136.61,130.48,128.57,128. 06,127.31,125.23,123.73,122.60,122.48,122.39,122.31,121.49,120.45,117.97,111.67,48.72,34.06,23.93.HRMS(ESI):found 521.2046,calcd for C 28 H 24 N8O3[M+H] + 521.2044

[0131] Example 29

[0132] Synthesis of N-(3-1,2,4-oxadiazol-3-yl)phenyl)-2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)acetamide

[0133] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with formic acid, resulting in a pale yellow solid, namely N-(3-1,2,4-oxadiazol-3-yl)phenyl)-2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ10.96(s,1H),8.03(s,1H),7.94–7.77(m,3H),7.62–7.50(m,2H),7.50–7.35(m,2H),7.02(s,2H),5.62(s,2H). 13 C NMR(151MHz,DMSO)δ166.31,156.47,141.95,141.78,139.85,138.71,136.52,130.9 1,127.64,125.20,124.15,123.72,122.21,120.42,118.97,112.21,111.62,48.67.

[0134] Example 30

[0135] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-methyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0136] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with acetic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-methyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO)δ10.91(s,1H),8.36(s,1H),8.10–7.79(m,2H),7.69(dd,J=17.1 ,9.4Hz,2H),7.59–7.33(m,3H),7.02(s,2H),5.63(s,2H),2.73(s,1H),2.64(s,2H). 13 C NMR (151MHz, DMSO) δ177.92,167.91,165.90,156.49,141.97,141.83,139.80,138.74,136.56,130.3 8,127.36,125.19,123.70,122.40,122.21,120.42,117.92,111.61,48.66,12.45.HRMS(ESI):found 417.1416,calcd for C 20 H 16 N8O3[M+H] + 417.1418

[0137] Example 31

[0138] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-ethyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0139] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with propionic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-ethyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ10.88(s,1H),8.37(s,1H),7.97–7.68(m,4H),7.62–7. 35(m,3H),7.05(s,2H),5.65(s,2H),3.14–2.87(m,1H),1.35(t,J=7.5Hz,3H).13 C NMR (151MHz, DMSO) δ181.73,167.79,165.91,156.49,141.97,141.82,139.82,138.74,136.56,130.38,1 27.41,125.20,123.70,122.41,122.19,120.42,117.90,111.61,48.66,20.03,10.88.HRMS(ESI):found 431.1580,calcd for C 21 H 18 N8O3[M+H] + 431.1575

[0140] Example 32

[0141] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-propyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0142] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with n-butyric acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-propyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (500MHz, DMSO-d6) δ10.82(s,1H),8.36(s,1H),7.92–7.84(m,2H),7.77–7.70(m,2H),7.52(t,J=7.9Hz,1H),7.47(t,J=7 .5Hz,1H),7.41(t,J=7.4Hz,1H),7.02(s,2H),5.64(s,2H),2.97(t,J=7.4Hz,2H),1.90–1.76(m,2H),0.98(t,J=7.4Hz,3H). 13 C NMR (126MHz, DMSO) δ180.79,167.83,165.95,156.52,142.00,141.85,139.86,138.76,136.59,130.41,127. 42,125.21,123.72,122.43,122.22,120.45,117.94,111.66,48.69,28.04,19.96,13.82.HRMS(ESI):found 445.17,calcd forC 22 H20 N8O3[M+H] + 445.17

[0143] Example 33

[0144] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-isopropyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0145] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with isobutyric acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-isopropyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO-d6)δ10.85(s,1H),8.33(t,J=1.7Hz,1H),7.97–7.68(m,5H),7.5 8–7.36(m,3H),7.02(s,2H),5.62(s,2H),3.34–3.28(m,1H),1.36(d,J=7.0Hz,6H). 13 C NMR (151MHz, DMSO) δ184.65,167.74,165.91,156.48,141.97,141.81,139.80,138.74,136.54,130.39,1 27.40,125.22,123.72,122.44,122.21,120.43,117.90,111.59,48.65,27.25,20.28.HRMS(ESI):found 443.1586,calcdfor C 22 H 20 N8O3[MH] - 443.1586

[0146] Example 34

[0147] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(2,3-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0148] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2,3-difluorobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(2,3-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ10.95(s,1H),8.03(s,1H),7.98–7.76(m,4H),7.76–7.51(m,3H),7.49–7.31(m,3H),7.02(s,2H),5.61(d,J=5.0Hz,2H). 13 C NMR (151MHz, DMSO) δ172.04,168.43,165.98,165.84,156.50,141.98,141. 82,139.96,139.56,138.74,136.57,130.53,130.04,126.86,126.47,126.3 6,126.34,125.20,123.70,122.86,122.74,122.62,122.57,122.19,121.78 ,120.42,117.99,116.74,114.31,114.26,111.62,48.69.HRMS(ESI):found 515.1383, calcd for C 25 H 16 F2N8O3[M+H] + 515.1386

[0149] Example 35

[0150] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(2,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0151] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2,4-difluorobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(2,4-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1H NMR (300MHz, DMSO-d6) δ10.89(s,1H),8.44(s,1H),8.35–8.20(m,1H),7.94–7.73(m,4H),7.69–7.32(m,6H),7.03(s,2H),5.62(d,J=13.0Hz,2H). 13 C NMR (151MHz, DMSO) δ172.25,168.29,165.96,156.50,141.98,141.82,139.92,138.74,136.57,133.31,133.23,130.49,126.97,125.20,12 3.70,122.61,122.50,120.42,117.99,113.77,113.60,111.63,109.32,106.59,106.41,106.24,48.69.HRMS(ESI):found515.1386,calcd for C 25 H 16 F2N8O3[M+H] + 515.1386

[0152] Example 36

[0153] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(2,5-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0154] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2,5-difluorobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(2,5-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO) δ10.99 (s, 1H), 8.22–7.69 (m, 6H), 7.67–7.35 (m, 5H), 7.06 (d, J = 16.8Hz, 2H), 5.64 (s, 2H). 13C NMR (151MHz, DMSO) δ166.31,165.66,156.47,141.97,141.95,141.84,141.78,139.85,138.74,138.71,136.55,1 36.52,130.91,129.17,127.65,125.21,124.16,123.73,122.22,120.43,119.12,118.97,112.21,111.62,48.66.

[0155] Example 37

[0156] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(2,6-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0157] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2,6-difluorobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(2,6-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ8.25(s,1H),7.78–7.58(m,5H),7.40(t,J=7.9Hz,1H),7.34–7.18(m,4H),6.87(s,2H),5.48(s,2H). 13 CNMR(151MHz,DMSO)δ168.86,168.84,168.81,168.46,165.98,161.43,161.40, 159.72,159.69,156.49,141.98,141.82,139.99,138.74,136.55,136.48,136.4 1,136.33,130.55,126.78,125.20,123.70,122.62,120.42,118.02,113.64,11 3.62,113.50,113.48,111.61,103.04,102.93,102.82,48.68.HRMS(ESI):found 513.1245,calcd forC 25 H 16 F2N8O3[MH] - 513.1241

[0158] Example 38

[0159] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(3,5-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0160] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 3,5-difluorobenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3,5-difluorophenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO) δ10.95(s,1H),8.03(s,1H),7.98–7.76(m,7H),7.76–7.51(m,1H),7.49–7.31(m,2H),7.02(s,2H),5.61(d,J=5.0Hz,2H). 13 C NMR (151MHz, DMSO) δ168.81,165.99,164.01,163.92,162.36,156.50,141.98,141.82,139.94,138.74,136.56,130.52,12 6.84,126.66,125.20,123.71,122.70,122.63,120.43,117.91,112.07,111.89,111.62,109.38,48.68.HRMS(ESI):found 515.1384,calcd for C 25 H 16 F2N8O3[M+H] + 515.1386

[0161] Example 39

[0162] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-benzyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0163] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with phenylacetic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-benzyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide.1 H NMR(300MHz,DMSO-d6)δ10.88(s,1H),8.33(s,1H),7.93–7.82(m,2H),7.7 9–7.67(m,2H),7.56–7.28(m,8H),7.04(s,2H),5.63(s,2H),4.43(s,2H). 13 CNMR(151MHz,DMSO)δ179.39,168.04,165.90,156.49,141.97,141.82,139.83,138.73,136.55,134.50,130.40,129. 53,129.24,127.81,127.20,125.19,123.70,122.43,122.29,120.41,117.92,111.61,48.66,32.45.HRMS(ESI):found 515.15,calcdfor C 26 H 20 N8O3[M+Na] + 515.15

[0164] Example 40

[0165] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(1-benzylethyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0166] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 2-phenylpropionic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(1-benzylethyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ10.89(s,1H),8.34(s,1H),7.88(t,J=8.8Hz,2H),7.82–7.68(m,2 H),7.56–7.27(m,8H),7.04(s,2H),5.63(s,2H),4.76–4.53(m,1H),1.72(d,J=7.1Hz,3H). 13C NMR (151MHz, DMSO) δ182.55,167.90,165.92,156.49,141.98,141.82,140.72,139.85,138.74,136.55,130.40, 129.34,127.92,127.82,127.23,125.19,123.70,122.46,122.29,120.42,117.91,111.62,48.67,37.88,20.04.

[0167] Example 41

[0168] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-phenylethyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0169] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 3-phenylpropionic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-phenylethyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR(300MHz,DMSO-d6)δ8.35(s,1H),7.98–7.66(m,4H),7.57–7.33(m,4H),7.32–7 .15(m,4H),7.03(s,2H),5.63(s,2H),3.31(t,J=7.6Hz,2H),3.11(t,J=7.6Hz,2H). 13 C NMR (151MHz, DMSO) δ180.17,167.78,165.92,156.49,141.97,141.83,140.10,139.83,138.74,136.55,130.39,128.87,1 28.75,127.30,126.84,125.20,123.71,122.42,122.26,120.42,117.94,111.60,48.66,32.08,27.99.HRMS(ESI):found 505.1746,calcd for C 27 H 22 N8O3[MH] - 505.1742

[0170] Example 42

[0171] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-butyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0172] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with n-valeric acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-butyl-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ10.87(s,1H),8.36(s,1H),7.88(t,J=9.2Hz,4H),7.58–7.35(m,3H),7.04(s,2 H),5.64(s,2H),2.99(t,J=7.5Hz,2H),1.76(p,J=7.5Hz,2H),1.42–1.32(m,2H),0.92(t,J=7.3Hz,3H). 13 C NMR (151MHz, DMSO) δ180.94,167.82,165.93,156.50,141.98,141.83,139.81,138.75,136.56,130.40 ,127.39,125.23,123.73,122.44,122.23,120.44,117.94,111.60,48.65,28.44,25.89,21.96,13.87.

[0173] Example 43

[0174] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(3-phenylpropyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0175] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 4-phenylbutyric acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3-phenylpropyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1H NMR (300MHz, DMSO-d6) δ11.09 (s, 1H), 8.39 (s, 1H), 7.88 (t, J = 7.5Hz, 2H), 7. 74(dd,J=18.0,7.9Hz,2H),7.50(dd,J=16.5,8.2Hz,2H),7.41(dd,J=12.0,4 .7Hz,1H),7.23(td,J=11.3,9.9,6.8Hz,5H),7.04(s,2H),5.67(s,2H),2.98 (t,J=7.4Hz,2H),2.68(q,J=7.2Hz,2H),2.07(td,J=17.5,16.3,8.9Hz,2H). 13 C NMR (151MHz, DMSO) δ180.57,167.82,165.91,156.49,141.98,141.87,141.41,139.89,138.74,136.51,130.30,128.81, 127.35,126.42,125.20,123.70,122.40,122.24,120.41,117.94,111.60,48.68,34.63,28.10,25.72.HRMS(ESI):found 521.2048,calcd for C 28 H 24 N8O3[M+H] + 521.2045

[0176] Example 44

[0177] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(4-phenylbutyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0178] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 5-phenylpentanoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(4-phenylbutyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1H NMR (300MHz, DMSO-d6) δ11.31(s,1H),8.40(s,1H),7.88(dd,J=7.9,3.5Hz,2H),7.79(d,J=7.9Hz,1H),7.69(d,J=7.5Hz,1H),7.47(dt,J=25.4,8. 5Hz,3H),7.22(dq,J=15.4,7.6Hz,5H),7.05(s,2H),5.69(s,2H),3.02(t ,J=7.2Hz,2H),2.62(s,2H),1.77(d,J=7.3Hz,2H),1.68(d,J=6.6Hz,2H). 13 C NMR (151MHz, DMSO) δ180.78,167.86,165.93,156.51,142.22,142.00,141.92,139.97,138.75,136.50,130.26,128.73,127 .32,126.17,125.21,123.71,122.36,122.26,120.42,117.93,111.62,48.70,34.96,30.59,26.02,25.97.HRMS(ESI):found 535.2199,calcd for C 29 H 26 N8O3[M+H] + 535.2201

[0179] Example 45

[0180] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(5-phenylpentyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0181] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with benzohexanoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(5-phenylpentyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1H NMR (300MHz, DMSO) δ11.58(s,1H),8.83(s,1H),8.38–8.26(m,2H),8.26–8.11(m,2H),8.03–7.80(m,3H),7.75–7. 55(m,5H),7.49(s,2H),6.12(s,2H),3.54–3.31(m,2H),2.35–2.15(m,2H),2.13–1.94(m,2H),1.90–1.74(m,2H). 13 C NMR (151MHz, DMSO) δ180.85,167.84,165.93,156.52,142.52,142.00,141.88,139.90,138.75,136.53,130.32,128.70,128.65,127 .37,126.06,125.21,123.72,122.42,122.27,120.42,117.95,111.59,48.66,35.33,30.81,28.38,26.19,26.15.HRMS(ESI):found 549.2362,calcd for C 30 H 28 N8O3[M+H] + 549.2358

[0182] Example 46

[0183] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(3-trifluoromethyl)phenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0184] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 3-trifluoromethylbenzoic acid, resulting in a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(3-trifluoromethyl)phenyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ10.09(s,1H),7.55(d,J=17.2Hz,2H),7.34–7.09(m,2H),7.09–6.80(m,5H),6.80–6.45(m,3H),6.15(s,2H),4.77(s,2H). 13C NMR (151MHz, DMSO) δ174.71,168.73,165.99,156.50,141.98,141.84,139.97,138.75,136.56,132.41,131.50,130.50,130.23,130.2 0,126.94,125.20,124.90,124.77,124.74,123.71,122.64,122.56,120.43,117.97,111.63,48.70.HRMS(ESI):found547.1449,calcd for C 26 H 17 F3N8O3[M+H] + 547.1449

[0185] Example 47

[0186] Synthesis of 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzi[d]imidazol-1-yl)-N-(3-(5-(4-phenylcyclohexyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide

[0187] Referring to the synthesis method of Example 2, the 3,4-difluorobenzoic acid in step 6 of Example 2 was replaced with 4-phenylcyclohexane-1-carboxylic acid, yielding a pale yellow solid, namely 2-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1H-benzo[d]imidazol-1-yl)-N-(3-(5-(4-phenylcyclohexyl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide. 1 H NMR (300MHz, DMSO-d6) δ11.03(s,1H),8.40(d,J=9.4Hz,1H),7.87(t,J=7.7Hz,2H),7.79–7.70(m,2H),7.58–7.35(m,3H), 7.34–7.23(m,3H),7.23–7.08(m,2H),7.04(s,2H),5.65(s,2H),3.25–3.07(m,1H),2.81–2.59(m,1H),2.36–1.50(m,8H). 13C NMR (151MHz, DMSO) δ183.48,167.77,165.94,156.51,147.10,142.00,141.87,139.90,138.76,136.56,130.36,128.80,127.42,127 .13,127.09,126.48,125.21,123.72,122.41,122.24,120.43,117.97,111.63,48.69,42.96,35.55,33.09,30.54.HRMS(ESI):found 561.2357,calcd forC 31 H 28 N8O3[M+H] + 561.2358

[0188] Table 1. Structural formulas of compounds in Examples 1-47

[0189]

[0190]

[0191]

[0192]

[0193]

[0194]

[0195]

[0196] The following are the pharmacodynamic tests and results of some compounds in this invention:

[0197] 1. Assay for butyrylcholinesterase inhibitory activity:

[0198] Reagents and reagents: Analytical compounds, AChE (EC 3.1.1.7, selected from electric eel), eqBChE (Ec 3.1.1.8, selected from horse serum). 5,5'-Dithiobis(2-nitrobenzoic acid) (DTNB), acetylthiocholine (ATC) iodide, and butyrylthiocholine (BTC) iodide were purchased from Sigma-Aldrich; tacrine hydrochloride (9-Amin-1,2,3,4-tetrahydroacridine hydrochloride hydrate) was purchased from BioTrend.

[0199] Instrument: THERMO Varioskan Flash full-wavelength multi-functional microplate reader.

[0200] Experimental methods:

[0201] (1) Prepare buffer solution: Dissolve 13.6g potassium dihydrogen phosphate in 1L of water, and adjust the pH to 8.0±0.1 with potassium hydroxide. Store the solution at 4℃ for later use.

[0202] (2) Preparation of 0.01M DTNB solution: Dissolve 0.396g DTNB and 0.15g sodium bicarbonate in 100mL of water to prepare 0.01M DTNB solution, store at -30℃ for later use.

[0203] (3) Preparation of 0.075M ATC and BTC solution: Dissolve 0.217g ATC in 10mL of water to prepare 0.075M ATC and BTC solution, store at -30℃ for later use; Dissolve 0.237g BTC in 10mL of water to prepare 0.075M BTC solution, store at -30℃ for later use.

[0204] (4) Preparation of AChE and BChE solutions: Dissolve 5000 units of AChE in 1 mL of 1% gel solution, then dilute with water to 100 mL to obtain an AChE solution with a concentration of 5 units / mL. Store at -30℃ for later use. Dissolve 5000 units of BChE in 1 mL of 1% gel solution, then dilute with water to 100 mL to obtain a BChE solution with a concentration of 5 units / mL. Store at -30℃ for later use.

[0205] (5) Preparation of test solution: Dissolve the test compound in DMSO to prepare a solution with a concentration of 10. -3 A solution of M was then diluted with methanol to obtain solutions with concentrations of 10. -4 10 -5 10 -6 10 -7 10 -8 10 -9 10 -10 A solution of M.

[0206] Before the experiment, all solutions were warmed to room temperature, and the AChE and BChE solutions were diluted with water by half to prepare an enzyme solution with a concentration of 2.5 units / mL. Background UV absorbance was measured using a blank buffer (3 mL). 100 μL of the test compound solution, 100 μL of DTNB solution, and 100 μL of enzyme solution were added to 3 mL of buffer. After adding 20 μL of BTC solution to trigger the reaction, timing was immediately started, and the test solution was rapidly mixed. UV absorbance was measured at 412 nM after 2 min. The blank control group was measured using an equal volume of methanol instead of the test compound solution. All tests were performed in triplicate. Using the UV absorbance of the blank control group as 100%, the absorbance values ​​of the test compound at various concentrations were recorded. The results were used to calculate the corresponding IC50 using GraphPad Prism™ software (GraphPad Software, San Diego, CA, USA) in a non-linear regression analysis model. 50 The values ​​are shown in Table 2.

[0207] All 47 compounds in Table 2 exhibited high selective inhibitory activity against BchE, with most drugs showing IC50 values. 50 Reaching the 10 nanomolar level.

[0208] Table 2. Results of Compound Target Activity Assay

[0209]

[0210]

[0211]

[0212] The inhibitory activity of each compound against the enzyme is indicated by "+", and "++" represents IC50. 50 The range is from 1 μM to 1 mM, and "+++" indicates IC. 50 The range is from 1 nM to 1 μM, and "++++" represents IC. 50 <1nM. b AChE (EC 3.1.1.7) from electric eel. c BuChE (EC 3.1.1.8) from horse serum.

[0213] The parent molecule of butyrylcholine ester inhibitor S06-1064 has the following structure:

[0214]

[0215] 2. L02 cytotoxicity assay:

[0216] Drugs and reagents: The test compound and DMEM medium (01-050-1A) were purchased from Biological Industries, FBS fetal bovine serum (04-001-1A) were purchased from Biological Industries, and MTT thiazolyl blue reagent (KGT525500) was purchased from Kaiji Biotechnology.

[0217] Instrument: THERMO Varioskan Flash full-wavelength multi-functional microplate reader.

[0218] Experimental methods:

[0219] (1) Cell seeding: The cells were seeded at a density of 5000 cells / well in a 96-well plate and incubated at 37°C in a 5% CO2 incubator for 24 hours.

[0220] (2) Discard the culture medium, wash once with PBS buffer, and add 100 μL of different concentrations of the compound prepared in DMEM culture medium to each well. The blank control group only added DMEM culture medium without the compound. Incubate the 96-well plate at 37°C in a 5% CO2 incubator for 24 h.

[0221] (3) Add 15 μL of MTT solution (5 mg / mL, i.e. 0.5% MTT) to each well in the dark and continue culturing for 2-3 h.

[0222] (4) Discard the culture medium, add 100 μL of dimethyl sulfoxide to each well, and gently shake to fully dissolve the crystals in the dimethyl sulfoxide. Measure the absorbance (OD) of each well at 490 nm using a multi-mode microplate reader. The OD value of the normal group was set as 100%. The cell viability was calculated using the following formula: Cell viability = (OD value of the drug-treated group - OD value of the blank group) / (OD value of the normal group - OD value of the blank group) × 100%. The experimental results are shown in Table 3. Cytotoxicity assessment of L02 hepatocytes confirmed that all compounds had excellent safety, with viability exceeding 50% at a concentration of 100 μM.

[0223] Table 3. Results of compound cytotoxicity assays

[0224]

[0225]

[0226]

[0227]

[0228] a. Survival rate, the survival rate (%) of L02 cells with a concentration of 100 μM of the compound, is indicated by "+".

[0229] ++++: ≥90% survival; +++: 75%–89% survival; ++: 50%–74% survival

[0230] 3. Nrf2 activation activity assay:

[0231] Experimental methods:

[0232] (1) In vitro cell culture. 293T cells transfected with the ARE luciferase reporter gene plasmid were cultured at 37°C and 5% CO2.

[0233] (2) 293T-ARE cells in the logarithmic growth phase were treated with 0.25% trypsin digestion solution to prepare a solution with a concentration of 4×10⁻⁶. 5 Cell suspension. Add 100 μL to each well of a 96-well microplate and incubate overnight.

[0234] (3) Prepare the test compound at twice the test concentration using culture medium, and add 100 μL to the corresponding wells. tert-butylhydroquinone (tBHQ) and dimethyl fumarate (DMF) are used as positive controls, and DMSO is used as a negative control. Incubate the 96-well microplate with the added compounds at 37°C and 5% CO2 for 12 h.

[0235] (4) Prepare the 5× lysis buffer in the luciferase detection kit as a 1× lysis buffer for later use.

[0236] (5) Remove the 96-well plate, aspirate the culture medium from the wells, wash the cells with 1×PBS buffer, and aspirate the PBS buffer after washing. Add 25 or 30 μL of 1× cell lysis buffer to each well and lyse on ice for 15 min. After lysis, let stand for 3-5 min, and aspirate 20 μL of supernatant to the corresponding 96-well white microplate.

[0237] (6) Place the white ELISA plate into the Thermo Scientific Luminoskan Asent chemiluminescence microplate reader. Before testing, add 100 μL of luciferase assay reagent to each well (prepared by uniformly mixing one vial of luciferase assay substrate and one vial of luciferase assay buffer from the luciferase assay kit). Take the reading within 1 minute after adding the assay reagent. The results are as follows: Figure 1 As shown.

[0238] Compared to the positive control drug dimethyl fumarate (DMF), several compounds exhibited superior Nrf2 activation effects. The induction folds of compounds 21, 28, 29, and 34 at 20 μM were comparable to or better than those of DMF at 20 μM, with compound 35 showing the highest induction fold. Among these, compounds 21, 34, and 35 stood out as leading drug candidates due to their potent BChE inhibition in the sub-10 nM range. Considering the susceptibility of the terminal methyl group in compound 21 to oxidation and potential metabolic instability, further pharmacological and mechanistic studies focused on compounds 34 and 35.

[0239] 4. ROS Measurement

[0240] Experimental reagent: Aβ 1–42 The following reagents were purchased from Biological Industries: Darmstadt (Germany), Reactive Oxygen Spectrometry Kit (Shanghai Beyotime Biotechnology Co., Ltd.), test compounds, DMEM medium (01-050-1A) purchased from Biological Industries, FBS fetal bovine serum (04-001-1A) purchased from Biological Industries, and MTT thiazolyl blue reagent (KGT525500) purchased from Kaiji Biotechnology.

[0241] Experimental consumables and instruments: 6-well plates, fluorescence inverted microscope

[0242] Experimental steps:

[0243] SH-SY5Y cells in the logarithmic growth phase were selected and cultured at a concentration of 2.5 × 10⁻⁶. 5 Cells were seeded at a density of 6-well plates and incubated at 37°C in a 5% CO2 incubator for 24 h. The culture medium was discarded, and the cells were washed once with PBS and pretreated for 24 h with either test compound 34 (10 μM) or 35 (10 μM). Subsequently, β-amyloid protein (Aβ) was added to the cell system. 1-42 The oligomer (5 μM) was incubated for another 24 hours. The culture medium was removed, and 1 mL of 10 μM DCFH-DA (diluted 1:1000) prepared with PBS was added to each well. The cells were incubated at 37°C for 30 min to ensure adequate contact between the probe and cells. The cell culture medium was removed, and the cells were washed three times with PBS to remove any uninfiltrated DAFH-DA. Visualization and evaluation were performed using a fluorescence microscope. The results are shown below. Figure 2 As shown.

[0244] Oxidative stress is characterized by the excessive and dysregulated production of reactive oxygen species (ROS). Under physiological conditions, ROS levels are maintained in a tightly controlled equilibrium and play a crucial role in various cellular signaling pathways. However, abnormal ROS accumulation disrupts redox homeostasis, ultimately leading to neuronal damage and impaired peripheral glial cells. Compared to the control group, Aβ...1-42 The significantly enhanced green fluorescence intensity in the treatment group confirmed the successful establishment of the ROS overproduction model. Both compounds (10 μM) significantly reduced ROS production, and neither showed significant anti-proliferative activity at the tested concentrations. These findings suggest that compounds 34 and 35 can effectively alleviate oxidative stress.

[0245] 5. Mouse behavioral studies

[0246] All animal experiments were approved by the Animal Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and the New Drug Evaluation Research Center of China Pharmaceutical University (Nanjing, China), and were strictly conducted in accordance with the U.S. National Institutes of Health's "Guidelines for the Care and Use of Laboratory Animals".

[0247] The recombinant human β-amyloid protein (Aβ) used in the experiment was pretreated with 1,1,1,3,3,3-hexafluoroisopropanol (HFIP). 1–42 The peptide was purchased from Merck Millipore (Darmstadt, Germany). Oligomeric Aβ 1–42 The preparation method followed previous literature and was used as a modeling stimulant. Mice were anesthetized with isoflurane, and oligomeric Aβ was injected bilaterally into the hippocampus via stereotactic surgery. 1–42 An Alzheimer's disease model was established. The experimental animals were divided into 9 groups (n=5 in each group): (1) Blank control group: intraperitoneal injection of solvent (ip); (2) Sham operation group: intravenous injection of saline (icv); (3) Model group: intravenous injection of oligomerized Aβ. 1–42 (10 μg / animal, icv); (4) Positive control group: intraventricular injection of oligomerized Aβ 1–42 (10 μg / animal, icv) combined with intraperitoneal injection of the positive control drug rivastigmine (1 mg / kg, ip); (5) Nrf2 activator 6 single drug group: intraventricular injection of oligomerized Aβ 1–42 (10 μg / animal, icv) combined with intraperitoneal injection of test compound 6 (1 mg / kg, ip); (6) BchE inhibitor 1064 monotherapy group: intraventricular injection of oligomerized Aβ 1–42 (10 μg / animal, icv) combined with intraperitoneal injection of test compound 1064 (1 mg / kg, ip); (7) Compound 6 + 1064 combination group: intraventricular injection of oligomerized Aβ 1–42 (10 μg / animal, icv) combined with intraperitoneal injection of test compound 6 (1 mg / kg, ip) and 1064 (1 mg / kg, ip); (8) Compound 34 group: intraventricular injection of oligomerized Aβ 1–42(10 μg / animal, icv) combined with intraperitoneal injection of test compound 34 (1 mg / kg, ip); (9) Compound 35 group: intraventricular injection of oligomerized Aβ 1–42 Compound 35 (1 mg / kg, ip) was administered via intraperitoneal injection in combination with 10 μg / animal. Compound 1064 is the parent molecule S06-1064 of the BChE inhibitor mentioned earlier, and compound 6 is the parent molecule of the Nrf2 activator, with the following structure:

[0248]

[0249] Behavioral testing includes:

[0250] Morris Water Maze (MWM) Test: Used to assess spatial learning and reference memory abilities. The experiment includes 5 days of orientation navigation training (a concealed platform is fixed in a quadrant of a pool with a diameter of 120 cm and a water depth of 40 cm, at a water temperature of 25°C). On the 6th day, the platform is removed and a 60-second exploration test is conducted, recording the percentage of time spent in the target quadrant and the number of times the original platform area is traversed.

[0251] Y-maze test: assesses working memory capacity. Mice were allowed to adapt for 5 minutes and then freely explore the maze for 5 minutes. The spontaneous alternation rate (%) and total number of arm entries were recorded.

[0252] New Object Recognition (NOR) Test: Conducted in two phases. First, train the user with two identical objects for 5 minutes to familiarize themselves with them. Then, replace the object with a new one and conduct a 5-minute test. Recognition and memory ability is expressed as a discrimination index: (Time spent exploring the new object - Time spent exploring the familiar object) / (Time spent exploring the new object + Time spent exploring the familiar object).

[0253] All behavioral processes were recorded using the Panlab SMART 3.0 video tracking system, and data analysis was performed using GraphPad Prism 5 software. The results are as follows: Figure 3 As shown. Injection of Aβ 1-42 Mice exhibited significant cognitive impairment in all tests (p<0.0001 vs. sham control): in the water maze (days 8–13), they displayed chaotic swimming patterns ( Figure 3 B) and a long escape latency period ( Figure 3 C) Demonstrates impaired spatial learning in mice; in the Y maze (day 14), mice showed reduced exploration of the new arm ( Figure 3 D and 3E) and the spontaneous alternation rate decreases ( Figure 3 F) indicates that the mice developed memory deficits; in the Novel Object Recognition (NOR) test (day 15), the discrimination index decreased ( Figure 3 G) and reduced time for exploring new objects ( Figure 3H) demonstrated impaired cognitive and memory abilities in mice. Single therapies of S06-1064 and 6 nearly improved these deficiencies, demonstrating the therapeutic potential of BChE inhibition and Nrf2 activation against Aβ-induced neurotoxicity. Notably, the combination therapy group exhibited superior cognitive improvement compared to either single-agent therapy, with dual-target molecules 34 and 35 showing the most significant effects, superior to single-agent therapy and drug combinations at equivalent doses (1 mg / kg). These dual-target molecules also offer additional advantages such as lower molecular weight, simplified dosage, and avoidance of potential drug interaction toxicities, collectively highlighting their therapeutic efficacy against AD enhancement.

Claims

1. A compound with a benzimidazole-aminofuran-oxadiazole skeleton or a pharmaceutically acceptable salt thereof, characterized in that, The structures of the compounds are shown in general formulas I and II: in, R 1 R 2 Each independently represents hydrogen or R 3 Represents hydrogen, methyl, or halogen; X represents carbon or nitrogen; n = 1, 2, 3; R 4 Represents hydrogen, C 1～6 Alkyl, substituted or unsubstituted phenyl, phenylalkyl; the substitution is selected from halogen, nitro, methoxy, C 1～6 Alkyl, halomethyl, cyclohexyl; the alkyl group in the phenylalkyl group is C10. 1～6 alkyl.

2. The compound of the benzimidazole-aminofuran-oxadiazole skeleton according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, R 1 R 2 Each independently represents hydrogen or R 3 Represents hydrogen, methyl, or bromine; X represents carbon or nitrogen; n = 1, 2; R 4 Represents hydrogen, C 1～6 Alkyl, substituted or unsubstituted phenyl, benzoalkyl; the substitution is selected from fluorine, chlorine, bromine, iodine, nitro, methoxy, C 1～6 Alkyl, trifluoromethyl, cyclohexyl; the alkyl group in the phenylalkyl group is C10. 1～6 alkyl.

3. The compound of the benzimidazole-aminofuran-oxadiazole skeleton according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, It is any of the following compounds or a pharmaceutically acceptable salt thereof:

4. The compound of the benzimidazole-aminofuran-oxadiazole skeleton according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The pharmaceutically acceptable salt is a salt formed by the compound and any one of the following acids: hydrochloric acid, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, malic acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, citric acid, benzenesulfonic acid, p-toluenesulfonic acid, or ferulic acid.

5. A method for preparing a compound of the benzimidazole-aminofuran-oxadiazole skeleton as described in any one of claims 1-4, characterized in that, The preparation method of the compound represented by general formula I includes the following steps:

6. The method for preparing the compound of the benzimidazole-aminofuran-oxadiazole skeleton or a pharmaceutically acceptable salt thereof according to claim 5, characterized in that, by Using the raw material as a starting material, it was dissolved in acetic acid / water solution under ice bath conditions. Sodium nitrite was added, and the mixture was stirred at room temperature to obtain an intermediate. The intermediate is then refluxed with hydroxylamine hydrochloride and potassium hydroxide in diethylene glycol dimethyl ether at 100-120°C to obtain the intermediate. by Using chloroacetyl chloride or chloropropionyl chloride and sodium bicarbonate as starting materials, the mixture is stirred in dichloromethane at room temperature to obtain an intermediate. Then with intermediates Cesium carbonate was refluxed in N,N'-dimethylformamide at 40-60°C to give an intermediate. The intermediate was then refluxed with hydroxylamine hydrochloride and potassium carbonate in anhydrous ethanol at 70-90°C to obtain the intermediate. Finally and The target compound was obtained by amide condensation. The above R 3 X,n is defined as described in claim 1.

7. A method for preparing a compound of the benzimidazole-aminofuran-oxadiazole skeleton as described in any one of claims 1-4, characterized in that, The preparation method of the compound represented by general formula II includes the following steps:

8. A method for preparing the compound of the benzimidazole-aminofuran-oxadiazole skeleton as described in claim 7, or a pharmaceutically acceptable salt thereof, characterized in that, by Using the raw material as a starting material, it was dissolved in acetic acid / water solution under ice bath conditions. Sodium nitrite was added, and the mixture was stirred at room temperature to obtain an intermediate. The intermediate is then refluxed with hydroxylamine hydrochloride and potassium hydroxide in diethylene glycol dimethyl ether at 100-120°C to obtain the intermediate. by Using chloroacetyl chloride and sodium bicarbonate as starting materials, the mixture was stirred in dichloromethane at room temperature to obtain an intermediate. intermediate The intermediate was obtained by reflux of cesium carbonate in N,N'-dimethylformamide at 40-60°C. The intermediate was then refluxed with hydroxylamine hydrochloride and potassium carbonate in anhydrous ethanol at 70-90°C to obtain the intermediate. Finally with R 4 The target compound was obtained by amide condensation with -COOH. The above R 4 The definition is as described in claim 1.

9. A pharmaceutical composition, characterized in that, Compounds comprising the benzimidazole-aminofuran-oxadiazole skeleton as described in any one of claims 1-4, or pharmaceutically acceptable salts thereof and pharmaceutically acceptable carriers thereof.

10. Use of a compound of the benzimidazole-aminofuran-oxadiazole skeleton as described in any one of claims 1-4, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for the prevention or treatment of Alzheimer's disease.

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