

Citation: Du Chuanqian, Xie Baohua, He Ming, Hu Zhiye, Liu Yu, He Xue, Liu Fanyu, Cheng Chen, Zhou Hai-Bing, Huang Shengtang, Dong Chun'e. Design, Synthesis and Biological Evaluation of Pyrano[2, 3-b]-naphthoquinone Derivatives as Acetylcholinesterase Inhibitors[J]. Chinese Journal of Organic Chemistry, 2020, 40(7): 2035-2044. doi: 10.6023/cjoc202002039

新型吡喃并[2, 3-b]萘醌类乙酰胆碱酯酶抑制剂的设计合成及生物活性研究
-
关键词:
- 吡喃并[2, 3-b]萘醌
- / 乙酰胆碱酯酶抑制剂
- / 阿尔兹海默症
- / 分子模拟
English
Design, Synthesis and Biological Evaluation of Pyrano[2, 3-b]-naphthoquinone Derivatives as Acetylcholinesterase Inhibitors
-
1. Introduction
Alzheimerʼs disease (AD), the most common form of irreversible dementia in the elderly, is a multi-factorial and progressive neurodegenerative disorder of the central nervous system clinically characterized by progressive decline in cognitive, executive and memory functions.[1-2] A study estimated that there were 50 million people worldwide living with AD in 2019 and this number will reach 138 million in 2050.[3-4] The etiology and pathogenesis of AD are not yet clear, but the studies have indicated that those may involve in several factors including tau hyperphosphorylation, oxidative stress, β-amyloid protein deposition and cholinergic injury.[5-6] Cholinergic hypothesis asserts that the extensive decrease of acetylcholine (ACh) in the nervous cells leads to cognitive and memory deficits in AD patients. In the past decades, the main strategy for the treatment of AD was focused on the amount of ACh in brain tissue based on cholinergic hypothesis.[7-8] Acetylcholinesterase (AChE), which plays an important role in the central and peripheral cholinergic nervous system, is an enzyme that catalyzes the hydrolysis of ACh and contributes to the formation of choline. In the cholinergic hypotheses above, the reduction of ACh is thought to be the root cause of memory and cognitive impairments or deficits, [9-11] the function of AChE is mostly recognized and has been extensively investigated. AChE inhibitors exert effects through inhibiting the AChE from breaking down ACh, thereby increasing both the level and duration of ACh. In various tissues of body, in addition to AChE, there is also another kind of cholinesterase, butyrylcholinesterase (BuChE). BuChE is mainly distributed in liver, plasma and cerebral white matter glial cells, which has little effect on central ACh levels in patients with mild to moderate AD. So, the poorly selective AChE inhibitors can inhibit the peripheral BuChE activity, which leads to peripheral ACh accumulation, and cause peripheral choline-like reactions such as nausea and vomiting.[12]
Currently, AChE inhibitors and NMDA receptor antagonists are widely used in the clinical treatment of AD. There are some AChE inhibitors including donepezil, rivastigmine, and tacrine (Scheme 1).[13] In particular, AChE inhibitors, such as donepezil, have become the most important drugs in clinic for the treatment of AD.[14] Although these drugs have good effects in clinical use, most of them only relieve symptoms and cannot completely cure AD, [15] thus their application is greatly limited. In clinical treatment of AD, the highly selective AChE inhibitor donepezil possessed a more favorable therapeutic index than tacrine, which could inhibit both of AChE and BuChE. As such, tacrine has been removed from the market due to the hepatotoxicity, significant drug-drug interactions, and serious side effects etc.[16] So far, the current development of AChE inhibitors mainly includes the following methods: (1) modification based on tacrine, (2) modification based on donepezil, (3) development of new structure AChE inhibitors.
Scheme 1
Although great progress in the strategy of modification of the skeleton based on tacrine and donepezil has been made, it was hindered in later studies due to poor biological activity and side effects.[17-21] Therefore, it is crucial to develop potent and highly selective AChE inhibitors with new structures. It has been reported that compounds containing a benzopyrone skeleton have good AChE inhibitory activity, such as compound 1, with IC50 value of 6.33 nmol/L.[22] As part of our long-term interest in neuro-protective research, we have devoted to the development of new AChE inhibitors. In previous studies, it has been found that pyrano[3, 2-c]chromene compounds exhibited good AChE inhibitory activity in vitro. In particular, compound 2, its inhibitory activity IC50 for AChE is 12.9 μmol/L.[23] Moreover, we found that the pyran structure played an important role in these compounds and many natural products, and their derivatives containing naphthoquinone moiety have multiple biological activities.[24] Based on these researches, we aimed to replace the chromene core structure with naphthoquinone in order to obtain series of piano[2, 3-b]naphthoquinone derivatives. Herein, we describe the design, synthesis and biological evaluation of pyrano[2, 3-b]naphthoquinone 3 as the AChE inhibitors. This work will not only provide a fundamental understanding of selective activity between AChE and BuChE, but also open new prospects on the design of highly selective AChE inhibitors.
2. Results and discussion
2.1 Chemistry
To investigate structure activity relationships (SARs) of these pyrano[2, 3-b]naphthoquinones as new class of AChE inhibitors, substituted benzene ring and other different sterically hindered substituents were introduced into the basic pyrano[2, 3-b]naphthoquinone skeleton in order to obtain a diverse of these compounds, which were accomplished in a stepwise fashion. Firstly, the substituted vinyldinitriles (5) were prepared using substituted aldehydes and malononitrile under microwave irradiation condition according to the known method.[25] By this means, we can easily and rapidly get intermediate in high yields. Then pyrano[2, 3-b]naphthoquinone derivatives 3a~3x were synthesized by Michael addition reaction from 2-hydroxy-1, 4- naphoquinone (4) and vinyldinitriles (5) with refluxing in anhydrous ethanol for 2 h in the presence of imidazole as shown in Scheme 2.[26] The desired products 3a~3x were afforded in good yields ranging from 80%~98%.
Scheme 2
2.2 Biological testing
The inhibitory activity profiles of pyrano[2, 3-b]naphthoquinone derivatives toward AChE enzymes (from the electric eel) and BuChE enzymes (from the equine serum) were evaluated by using the spectro- photometric Ellman assay, [27] in which AChE inhibitor donepezil was used as reference drug for comparison. With the AChE inhibition rate at 20 and 50 μmol/L, IC50 for AChE and BuChE of all compounds were summarized in Table 1. As shown in Table 1, it can be seen that most compounds had good inhibitory activities to AChE at 50 μmol/L, especially compounds 3f (Entry 6), 3g (Entry 7), 3h (Entry 8), 3i (Entry 9), 3k (Entry 11), 3l (Entry 12) and 3m (Entry 13), whose inhibitory activities were more than 80%. When the concentration of the inhibitors is at 20 μmol/L, most compounds showed over half inhibitory efficacy. Compounds 3f, 3k and 3q still displayed significant AChE inhibitory activities which were even more than 80%.
表 1
Entry Compd. AChE inhibition/% IC50c/(μmol•L-1) 20 μmol/La 50 μmol/Lb AChE BuChE 1 3a 59.66 67.87 10.40±0.90 >200 2 3b 55.94 67.99 6.09±0.50 >200 3 3c 60.25 69.12 2.37±0.30 >200 4 3d 51.14 73.52 20.70±1.80 >200 5 3e 63.90 74.83 4.31±0.10 >200 6 3f 80.80 87.59 3.67±0.10 >200 7 3g 67.79 86.52 6.03±0.10 >200 8 3h 68.46 89.61 6.50±0.08 >200 9 3i 68.80 82.38 12.70±0.40 >200 10 3j 32.35 62.70 37.10±2.50 >200 11 3k 80.32 86.54 3.82±0.20 >200 12 3l 67.22 81.77 11.80±1.27 >200 13 3m 66.68 80.63 4.17±0.59 >200 14 3n 70.90 74.91 1.22±0.12 >200 15 3o 57.17 64.96 2.25±0.06 >200 16 3p 63.83 71.41 5.29±0.02 >200 17 3q 80.15 78.78 4.36±1.09 >200 18 3r 22.70 27.54 >50 >200 19 3s 61.64 67.57 4.43±0.91 >200 20 3t 59.63 63.27 19.40±0.42 >200 21 3u 35.89 65.47 22.10±1.98 >200 22 3v 36.33 43.51 >50 >200 23 3w 35.89 65.47 25.50±2.26 >200 24 3x 20.99 34.47 >100 >200 25 Donepezil — — 0.03±0.001 29.7±2.46 a Data are represented as AChE inhibition (%) at 20 μmol/L. b Data are represented as AChE inhibition (%) at 50 μmol/L. c Data are represented as mean±SD. All experiments were independently carried out at least three times. Among the target compounds, the unsubstituted pyrano[2, 3-b]naphthoquinone derivative 3a exhibited an IC50 value of (10.40±0.90) μmol/L (Entry 1) towards AChE and was less potent than reference agent donepezil [AChE IC50=(0.03±0.001) μmol/L]. Based on the structure of compound 3a, we have modified the structure of the benzene ring and replaced the benzene ring with other hetero-cyclic groups. We found that compounds containing benzene ring had better inhibitory activity, especially compound 3n, with IC50 value up to (1.22±0.12) μmol/L, which was the best among all the synthesized compounds. At the same time, the substituents at different positions on the phenyl ring have little effect on inhibitory activity. Focusing on the halogen group, chlorine-substituted derivatives showed much better activity than bromine-substituted ones. In comparison of compounds 3f with 3g, 3k with 3m, 3r with 3s, it is found that electron-withdrawing or electron-donating groups have little effect on the inhibitory activity of this type of compounds. Compared with the compounds 3u~3x, we could see that the inhibitory activity of these compounds on AChE was significantly reduced, which indicated that the larger the substituent of the pyrano[2, 3-b]naphthoquinone, the weaker its inhibitory activity. Especially, compound 3x with the maximum steric hindrance has an IC50 value more than 100 μmol/L. The structure activity relationship (SAR) is shown in Scheme 3. The inhibitory activities of pyrano[2, 3-b]naphthoquinone derivatives toward BuChE are also showed in Table 1. Surprisingly, most compounds exhibited lower inhibitory activity against BuChE even at 200 μmol/L. The results indicate that these compounds possess much higher selectivity to AChE than BuChE.
Scheme 3
The CLogP values of these synthesized compounds were predicted. To our delight, the CLogP values of the most compounds were between 2 and 4. Additionally, compound 3n exhibited a suitable CLogP value (2.66), consistent with our drug-like physical criteria. However, replacement of the phenyl moiety with heterocyclic aromatic rings, such as indole and benzo[1, 3]dioxole resulted in a dramatic increase of CLogP values.
Taken together, compound 3n, as a novel type of acetylcholinesterase inhibitor, has shown potential as a lead for development of drugs for Alzheimer's Disease. According to the results of SARs, molecular docking (see below for details) of these pyrano[2, 3-b]naphthoquinones and the structures of inhibitors reported in literatures, further optimization of compound 3n could be focused on the modification of naphthoquinone core structure by introducing different substituents, [24, 26] installation of a side chain of donepezil or similar one through the NH2 or CN group of 3n to increase the interaction with acetylcholinesterase, [28-30] or using compound 3n to react with cyclohexanone directly to obtain five-membered ring fused analogues to increase the structural diversity of this type of inhibitors.[31-32]
The mechanism of inhibition of AChE was investigated using compound 3n, one of the most potent AChE inhibitors. The Lineweaver-Burk double reciprocal plots were constructed with a range of substrate concentrations in Figure 1. According to the result, the obtained double reciprocal (Lineweaver-Burk) plot stands for a mixed-type inhibition pattern for compound 3n.
图 1
To explore the possible binding modes and highly selective inhibition of compound 3n and donepezil towards AChE, the docking experiments were carried out based on the published crystal structure of the AChE protein (PDB:6O4W) by using AutoDock 4.2 and UCSF Chimera (Figure 2). Figure 2A showed the docking result of the reference drug donepezil with the AChE protein displayed by 3D plots. We could see that donepezil has a slender chain structure, which can well occupy the binding pocket of the protein, especially the basic side chain located in the hydrophilic part of the binding pocket. The oxygen atom in the methoxyl group can form two hydrogen bonds with H2O 832, and the lengths of the hydrogen bonds were both 0.301 nm. The carbonyl oxygen atom could form a hydrogen bond with Phe295, and the length of the hydrogen bond was 0.290 nm. In addition, the benzene rings of Trp286 and Tyr337 have π-π stacking interactions with the benzene rings in donepezil, respectively. These effects in donepezil result in good inhibitory potency towards AChE. Although CLogP of donepezil is high (CLogP=4.73), but its basic side chain might compensate this shortcoming and make donepezil have good hydrophilic interaction with binding pocket of the protein.
图 2
Then we studied the binding model of compound 3n with AChE. As shown in Figure 2B, compound 3n could bind to the same binding pocket of the AChE protein compared with donepezil. It was found that the oxygen in the carbonyl group of compound 3n could form a 0.279 nm hydrogen bond with Ser122, and also had moderate interaction with Ser200, Phe288. In addition, the CN group on the 3-position of the benzene ring occupied most of the binding pocket of the protein and formed a hydrogen bond with Ser200. Because the C≡N bond length was longer than other halogen substituents, it resulted in a closer distance between the CN group and the amino acid residue Ser200 than other substituents. This might be the reason why compound 3n had better inhibitory activity towards AChE than other compounds. Also, the low CLogP of compound 3n may contribute to its good inhibitory activity. However, we could see that due to the shorter diameter of compound 3n, only part of the binding pocket could be filled when compared to donepezil, resulting in less inhibitory activity of this series of compounds compared with donepezil.
To further understand the high selectivity for AChE of compound 3n, we studied the interaction between donepezil, compound 3n and AChE protein displayed by 2D plots (Figures 2C and 2D). It is noteworthy that the hydrogen bond and π-π stacking interaction are as same as those shown in 3D plots. These results may explain the high selectivity for AChE of these compounds.
3. Conclusions
In conclusion, based on a structure-diversity oriented drug design strategy, in this study we have integrated the basic skeleton of naphthoquinone pharmacophore with the bioactive anti-AChE motifs of pyrano[3, 2-c]chromene structure we obtained previously, and synthesized 24 novel pyrano[2, 3-b]1, 4-naphthoquinone derivatives. In the preliminary biological test, most of these compounds showed good inhibitory activities for AChE and no BuChE inhibition activity, inticating that these compounds had extremely high selectivity. Among these synthesized compounds, compound 3n was significantly potent and highly selective for AChE, with an IC50 value of 1.22 μmol/L and displayed the mixed type of inhibition in vitro. At the same time, the activity difference of these compounds and donepezil was also investigated. It was found that in most cases the compounds with better inhibitory activity had lower CLogP. Molecular modeling results point out that compound 3n could bind to the same binding pocket of the AChE protein compared with donepezil but had no binding function with BuChE protein. These new compounds can be considered as interesting new chemical entities with potential therapeutic application for Alzheimer's disease.
4. Experimental section
4.1 Materials and methods
NMR spectra were recorded at 400 MHz for 1H NMR spectra and 100 MHz for 13C NMR spectra at room temperature by Bruker Biospin AV400 and calibrated from residual solvent signals. The residual solvent peak was used as an internal reference: proton (DMSO-d6, δ=2.50, H2O δ=3.33), carbon (DMSO-d6, δ=39.52). Mass spectra (MS) were recorded on an IonSpec 4.7 Tesla FTMS using the DART Positive. All microwave irradiation experiments were carried out using a microwave oven XH-100B from Xianghu Company, China. The temperature in the MW experiments was measured by an eternal IR sensor. Purification by chromatography was performed using 230~400 mesh SiO2 with compressed air as a source of positive pressure. Melting points were determined on a Thermo Scientific electrothermal digital melting point apparatus and were uncorrected. Thin-layer chromatography (TLC) was performed on silica gel aluminium sheets with an F-254 indicator. Visualization was accomplished by UV light. Analytical-grade solvents were purchased, and used without further purification.
4.2 Chemistry
4.2.1 General procedure for the synthesis of 3
A mixture of the substituted aldehydes (2 mmol) and malononitrile (2 mmol) in anhydrous EtOH (10 mL) was subjected to microwave irradiation condition (60 ℃, 500 W) for 30 min. The progress of the reaction was monitored by thin-layer chromatography (TLC). The mixture was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate (50 mL), and then concentrated. The residue was purified by column chromatography to afford 5 as solid [V(petroleum ether):V(ethyl acetate)=6:1]. The characterization data for intermediates 5a~5g have been reported.[33-36]
Anhydrous EtOH (20 mL) was added to the solid mixture of the substituted intermediate 5 (4 mmol), 2-hydroxy-1, 4-naphoquinone 4 (4 mmol) and imidazole (0.8 mmol) in vacuo. The resulting solution was stirred and heated to reflux (90 ℃) for 2 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was gradually cooled to room temperature and the solvent was concentrated by rotary vacuum evaporator. 10 mL of water was added to the residue and the mixture was extracted with ethyl acetate (20 mL×3). Then the organic layer was washed with brine and dried with anhydrous sodium sulfate. Evaporation of ethyl acetate gave a brown to red residue, which were purified by column chromatography with dichloromethane to obtain final compounds 3 [V(CH2Cl2):V(MeOH)=200:1].
2-Amino-5, 10-dioxo-4-phenyl-5, 10-dihydro-4H-benzo-[g]chromene-3-carbonitrile (3a): Orange solid, 96% yield. m.p. 304~306 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.05 (dd, J=6.2, 2.4 Hz, 1H), 7.93~7.79 (m, 3H), 7.36 (s, 2H), 7.32 (d, J=4.1 Hz, 4H), 7.23 (dt, J=8.6, 4.2 Hz, 1H), 4.61 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.3, 158.8, 149.4, 144.0, 134.9, 134.6, 131., 131.1, 129.0, 128.1, 127.5, 126.5, 126.2, 122.4, 119.8, 57.9, 36.9; HRMS calcd for C20H12N2O3Na [M+Na]+ 351.0740, found 351.0753.
2-Amino-4-(4-fluorophenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3b): Yellow solid, 93% yield. m.p. 293~295 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.09~8.02 (m, 1H), 7.85 (dd, J=6.7, 5.3 Hz, 3H), 7.38 (t, J=6.8 Hz, 4H), 7.13 (t, J=8.8 Hz, 2H), 4.65 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.3, 162.7 (d, JFC=242.4 Hz), 158.7, 149.4, 140.3, 134.9, 134.6, 131.4, 131.1, 130.2 (d, JFC=8.2 Hz), 126.5, 126.2, 122.1, 119.7, 115.7 (d, JFC=21.8 Hz), 57.7, 36.2; HRMS calcd for C20H11FN2O3Na [M+Na]+ 369.0646, found 369.0632.
2-Amino-4-(4-chlorophenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3c): Orange solid, 88% yield. m.p. 248~250 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.05 (d, J=6.3 Hz, 1H), 7.85 (dd, J=10.2, 7.2 Hz, 3H), 7.40 (s, 2H), 7.37 (s, 4H), 4.64 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.2, 158.7, 149.5, 143.1, 134.9, 134.6, 132.1, 131.4, 131.1, 130.1, 128.9, 126.5, 126.2, 121.8, 119.6, 57.4, 36.4; HRMS calcd for C20H11ClN2O3Na [M+Na]+ 385.0350, found 385.0346.
2-Amino-4-(4-bromophenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3d): Orange solid, 91% yield. m.p. 287~289 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.05 (d, J=6.1 Hz, 1H), 7.86 (d, J=8.0 Hz, 3H), 7.51 (d, J=8.2 Hz, 2H), 7.41 (s, 2H), 7.31 (d, J=8.3 Hz, 2H), 4.63 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.2, 158.7, 149.5, 143.5, 134.9, 134.6, 131.8, 131.4, 131.1, 130.5, 126.5, 126.2, 121.7, 120.6, 57.4, 55.3, 36.5; HRMS calcd for C20H11BrN2O3Na [M+Na]+ 428.9845, found 428.9839.
2-Amino-5, 10-dioxo-4-(p-tolyl)-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3e): Orange solid, 92% yield. m.p. 243~245 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.04 (d, J=6.2 Hz, 1H), 7.90~7.78 (m, 3H), 7.33 (s, 2H), 7.19 (d, J=7.9 Hz, 2H), 7.11 (d, J=7.9 Hz, 2H), 4.56 (s, 1H), 2.25 (s, 3H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.3, 158.7, 149.2, 141.1, 136.7, 134.9, 134.5, 131.4, 131.0, 129.6, 128.0, 126.5, 126.2, 122.5, 119.8, 57.9, 55.3, 36.5, 21.0; HRMS calcd for C21H14N2O3Na [M+Na]+ 365.0897, found 365.0890.
2-Amino-4-(4-methoxyphenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3f): Yellow solid, 98% yield. m.p. 247~249 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.04 (d, J=5.3 Hz, 1H), 7.85 (d, J=11.1 Hz, 3H), 7.32 (s, 2H), 7.23 (d, J=8.2 Hz, 2H), 6.86 (d, J=8.2 Hz, 2H), 4.56 (s, 1H), 3.71 (s, 3H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.3, 158.7, 149.0, 136.1, 134.9, 134.5, 131.4, 131.0, 129.3, 126.5, 126.2, 122.6, 119.9, 114.3, 58.1, 55.5, 36.1; HRMS calcd for C21H14N2O4Na [M+Na]+ 381.0846, found 381.0840.
2-Amino-4-(4-nitrophenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3g): Brown solid, 90% yield. m.p. 241~243 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.18 (d, J=8.6 Hz, 2H), 8.07 (d, J=7.6 Hz, 1H), 7.89~7.82 (m, 3H), 7.66 (d, J=8.6 Hz, 2H), 7.51 (s, 2H), 4.82 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.2, 158.8, 151.5, 149.9, 146.9, 135.0, 134.7, 131.3, 131.1, 129.6, 126.5, 126.2, 124.2, 121.0, 119.4, 110.0, 56.7, 36.9; HRMS calcd for C20H11N3O5Na [M+Na]+ 396.0591, found 396.0582.
2-Amino-4-(3-chlorophenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3h): Orange solid, 88% yield. m.p. 257~259 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.06 (d, J=7.9 Hz, 1H), 7.90~7.82 (m, 3H), 7.42 (s, 3H), 7.32 (dt, J=11.4, 7.5 Hz, 3H), 4.66 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.2, 158.7, 149.7, 146.5, 134.9, 134.6, 133.6, 131.4, 131.2, 130.8, 128.0, 127.5, 127.0, 126.5, 126.2, 121.3, 119.6, 57.4, 36.7; HRMS calcd for C20H11ClN2O3Na [M+Na]+ 385.0350, found 385.0345.
2-Amino-5, 10-dioxo-4-(m-tolyl)-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3j): Orange solid, 94% yield. m.p. 251~253 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.06 (d, J=8.4 Hz, 1H), 7.95~7.77 (m, 3H), 7.34 (s, 2H), 7.20 (t, J=7.5 Hz, 1H), 7.11 (d, J=10.9 Hz, 2H), 7.04 (d, J=7.3 Hz, 1H), 4.56 (s, 1H), 2.27 (s, 3H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.3, 158.7, 149.3, 144.0, 138.2, 134.9, 134.6, 131.4, 131.1, 128.9, 128.5, 128.2, 126.5, 126.2, 125.34 122.4, 119.8, 58.0, 36.9, 21.4; HRMS calcd for C21H14N2O3Na [M+Na]+ 365.0897, found 365.0883.
2-Amino-4-(3-methoxyphenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3k): Yellow solid, 87% yield. m.p. 245~247 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.05 (dd, J=6.8, 1.9 Hz, 1H), 7.86 (dd, J=6.9, 6.3 Hz, 3H), 7.35 (s, 2H), 7.23 (t, J=7.9 Hz, 1H), 6.93~6.84 (m, 2H), 6.81 (d, J=8.1 Hz, 1H), 4.59 (s, 1H), 3.73 (s, 3H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.3, 159.8, 158.8, 149.4, 145.6, 134.9, 134.5, 131.4, 131.1, 130.1, 126.5, 126.2, 122.2, 120.3, 119.8, 114.3, 112.4, 57.8, 55.4, 36.8; HRMS calcd for C21H14N2O4Na [M+Na]+ 381.0846, found 381.0841.
2-Amino-4-(3-hydroxyphenyl)-5, 10-dioxo-5, 10-dihydro- 4H-benzo[g]chromene-3-carbonitrile (3l): Orange solid, 90% yield. m.p. 244~246 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 9.43 (s, 1H), 8.04 (s, 1H), 7.87 (d, J=8.4 Hz, 3H), 7.35 (s, 2H), 7.10 (t, J=7.7 Hz, 1H), 6.82~6.66 (m, 2H), 6.62 (d, J=7.5 Hz, 1H), 4.51 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 182.9, 177.3, 158.9, 157.9, 149.1, 145.3, 135.0, 134.6, 131.4, 131.0, 130.0, 126.5, 126.3, 122.7, 119.8, 118.7, 114.9, 114.6, 57.8, 36.7; HRMS calcd for C20H12N2O4Na [M+Na]+ 367.0689, found 367.0683.
2-Amino-4-(3-nitrophenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3m): Yellow solid, 86% yield. m.p. 287~289 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.07 (d, J=7.6 Hz, 1H), 7.86 (dd, J=11.7, 9.0 Hz, 4H), 7.72 (t, J=6.9 Hz, 2H), 7.55 (t, J=7.8 Hz, 1H), 7.44 (s, 2H), 4.74 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 183.1, 177.2, 158.7, 150.0, 145.8, 134.9, 134.6, 133.3, 131.8, 131.4, 131.3, 131.2, 130.1, 126.4, 126.2, 120.8, 119.5, 119.1, 112.0, 57.2, 36.6; HRMS calcd for C20H11N3O5Na [M+Na]+ 396.0591, found 396.0585.
2-Amino-4-(3-cyanophenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3n): Red solid, 87% yield. m.p. 281~283 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.21 (s, 1H), 8.10~8.02 (m, 2H), 7.86 (s, 2H), 7.72 (t, J=7.1 Hz, 1H), 7.63 (t, J=7.9 Hz, 1H), 7.52 (s, 2H), 7.46 (s, 1H), 4.88 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 182.9, 177.1, 159.7, 158.7, 149.3, 145.4, 134.8, 134.4, 131.3, 131.0, 130.0, 126.4, 126.1, 122.0, 120.2, 119.7, 117.3, 114.1, 112.3, 57.7, 36.7; HRMS calcd for C21H11- N3O3Na [M+Na]+ 376.0693, found 376.0689.
2-Amino-4-(2-chlorophenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3o): Red solid, 86% yield. m.p. 308~310 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.08 (d, J=7.4 Hz, 1H), 7.85 (d, J=4.7 Hz, 3H), 7.43 (dd, J=9.3, 6.9 Hz, 2H), 7.38 (s, 2H), 7.30~7.20 (m, 2H), 5.16 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 182.9, 177.2, 158.8, 150.0, 141.4, 135.0, 134.6, 132.4, 131.3, 131.0, 130.9, 129.8, 129.1, 128.3, 126.5, 126.2, 121.7, 119.3, 56.6, 33.9; HRMS calcd for C20H11ClN2O3Na [M+Na]+ 385.0350, found 385.0346.
2-Amino-4-(2-bromophenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3p): Red solid, 81% yield. m.p. 272~274 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.07 (d, J=7.3 Hz, 1H), 7.89~7.82 (m, 3H), 7.60 (d, J=7.9 Hz, 1H), 7.45~7.34 (m, 3H), 7.30 (t, J=7.4 Hz, 1H), 7.16 (t, J=7.5 Hz, 1H), 5.16 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 182.9, 177.3, 158.7, 149.9, 143.2, 135.0, 134.6, 133.0, 131.4, 131.0, 131.0, 129.4, 128.9, 126.5, 126.2, 123.1, 121.8, 119.2, 56.8, 36.2; HRMS calcd for C20H11BrN2O3Na [M+Na]+ 428.9845, found 428.9831.
2-Amino-5, 10-dioxo-4-(o-tolyl)-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3q): Orange solid, 92% yield. m.p. 257~259 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.06 (d, J=7.2 Hz, 1H), 7.84 (d, J=5.1 Hz, 3H), 7.31 (s, 2H), 7.20~7.13 (m, 2H), 7.13~7.06 (m, 2H), 4.89 (s, 1H), 2.58 (s, 3H); 13C NMR (100 MHz, DMSO-d6) δ: 183.1, 177.3, 158.5, 149.5, 143.1, 135.2, 135.0, 134.6, 131.3, 131.0, 130.5, 128.6, 127.2, 126.5, 126.2, 123.3, 119.8, 57.8, 32.4, 19.6; HRMS calcd for C21H14N2O3Na [M+Na]+ 365.0897, found 365.0894.
2-Amino-4-(2-methoxyphenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitril (3r): Orange solid, 83% yield. m.p. 259~261 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.06 (d, J=5.4 Hz, 1H), 7.85 (d, J=6.2 Hz, 3H), 7.20 (d, J=9.2 Hz, 4H), 7.00 (d, J=8.5 Hz, 1H), 6.86 (t, J=7.4 Hz, 1H), 4.93 (s, 1H), 3.78 (s, 3H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.4, 159.3, 157.2, 150.0, 135.0, 134.5, 131.7, 131.4, 130.9, 129.5, 128.8, 126.5, 126.2, 122.3, 121.1, 119.9, 112.1, 56.9, 56.2, 31.5; HRMS calcd for C21H14N2O4Na [M+Na]+ 381.0846, found 381.0839.
2-Amino-4-(2-nitrophenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3s): Orange solid, 88% yield. m.p. 244~245 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.05 (d, J=6.6 Hz, 1H), 7.93 (d, J=7.9 Hz, 1H), 7.83 (s, 3H), 7.69~7.58 (m, 2H), 7.58~7.40 (m, 3H), 5.40 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 183.1, 177.2, 159.3, 149.5, 149.0, 138.3, 135.0, 134.7, 134.2, 131.8, 131.2, 131.0, 128.8, 126.5, 126.3, 124.5, 121.6, 56.1, 55.3, 31.5; HRMS calcd for C20H11N3O5Na [M+Na]+ 396.0591, found 396.0585.
2-Amino-4-(4-bromo-2-fluorophenyl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3t): Yellow solid, 85% yield. m.p. 260~262 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.07 (d, J=7.8 Hz, 1H), 7.86 (d, J=7.5 Hz, 3H), 7.54 (d, J=9.8 Hz, 1H), 7.50~7.30 (m, 4H), 4.91 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 182.9, 177.2, 163.6 (d, JFC=236.3 Hz), 159.0, 150.0, 135.0, 134.9 (d, JFC=37.2 Hz), 132.4, 131.3, 131.0, 130.5, 128.4, 126.5, 126.4 (d, JFC=29.2 Hz), 120.7, 119.4, 119.3, 119.1, 55.9, 55.3, 30.5; HRMS calcd for C20H10BrFN2O3Na [M+Na]+ 446.8751, found 446.8766.
2-Amino-4-cyclohexyl-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3u): Yellow solid, 89% yield. m.p. 248~250 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.01 (t, J=8.3 Hz, 2H), 7.90~7.83 (m, 2H), 7.32 (s, 2H), 1.53 (dd, J=12.4, 11.1 Hz, 7H), 1.31 (dd, J=13.0, 11.2 Hz, 1H), 1.18~1.00 (m, 3H), 0.88 (dd, J=12.1, 8.2 Hz, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 183.4, 177.1, 161.6, 150.5, 134.9, 134.5, 131.68, 131.1, 126.4, 124.1, 121.2, 52.2, 44.5, 36.3, 30.7, 27.5, 26.6, 26.3, 26.0; HRMS calcd for C20H18N2O3Na [M+Na]+ 357.1210, found 357.1205.
2-Amino-4-(benzo[d][1, 3]dioxol-4-yl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3v): Yellow solid, 91% yield. m.p. 263~265 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.10~8.00 (m, 1H), 7.87 (dd, J=10.2, 8.5 Hz, 3H), 7.35 (s, 2H), 6.89~6.72 (m, 3H), 6.04 (s, 1H), 5.93 (s, 1H), 4.72 (s, 1H); 13C NMR (100 MHz, DMSO-d6) δ: 182.88, 177.3, 159.1, 149.5, 147.6, 145.3, 135.1, 134.6, 131.4, 130.9, 126.5, 126.3, 125.3, 122.1, 122.1, 121.0, 119.7, 107.9, 101.4, 55.9, 32.1; HRMS calcd for C21H12- N2O5Na [M+Na]+ 395.0638, found 395.0627.
tert-Butyl3-(2-amino-3-cyano-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromen-4-yl)-1H-indole-1-carboxylat (3w): Yellow solid, 83% yield. m.p. 265~267 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 8.04 (dd, J=11.6, 7.7 Hz, 2H), 7.89~7.80 (m, 3H), 7.69 (s, 1H), 7.63 (d, J=7.8 Hz, 1H), 7.40 (s, 2H), 7.33 (t, J=7.7 Hz, 1H), 7.26 (t, J=7.5 Hz, 1H), 5.01 (s, 1H), 1.62 (s, 9H); 13C NMR (100 MHz, DMSO-d6) δ: 183.0, 177.3, 159.1, 149.4, 149.4, 135.2, 135.0, 134.6, 131.5, 131.1, 128.9, 126.5, 126.2, 124.9, 124.8, 123.2, 123.2, 121.4, 119.9, 119.7, 115.4, 84.5, 56.7, 28.1, 27.9; HRMS calcd for C27H21N3O5Na [M+Na]+ 490.1373, found 490.1369.
2-Amino-4-(1-(4-methoxybenzoyl)-1H-indol-3-yl)-5, 10-dioxo-5, 10-dihydro-4H-benzo[g]chromene-3-carbonitrile (3x): Yellow solid, 80% yield. m.p. 257~259 ℃; 1H NMR (400 MHz, DMSO-d6) δ: 9.43 (s, 2H), 8.28 (s, 1H), 7.90 (dd, J=5.1, 3.1 Hz, 1H), 7.87 (dd, J=5.2, 3.0 Hz, 1H), 7.80~7.75 (m, 2H), 7.71 (dd, J=14.6, 7.5 Hz, 2H), 7.65~7.56 (m, 2H), 7.46 (dd, J=10.9, 3.0 Hz, 1H), 7.12 (d, J=5.0 Hz, 2H), 6.85 (dd, J=5.0, 3.0 Hz, 1H), 4.29 (s, 1H), 3.79 (s, 3H); 13C NMR (100 MHz, DMSO-d6) δ: 183.5, 176.2, 166.3, 160.1, 157.4, 154.1, 135.4, 134.5, 134.3, 131.9, 131.4, 130.2, 128.6, 128.1, 127.1, 127.0, 126.2, 125.6, 124.3, 122.7, 122.4, 119.5, 116.6, 116.0, 113.6, 56.0, 55.3, 28.4; HRMS calcd for C30H19N3O5Na [M+Na]+ 524.1217, found 524.1213.
4.3 Biological assays
4.3.1 Procedure for the anti-AChE and BuChE activity assays
The inhibition ability of pyrano[2, 3-b]1, 4-naphthoquin- one derivatives against acetylcholinesterase (AChE, E.C. 3.1.1.7, from the electric eel) and butyrylcholinesterase (BuChE, E.C. 3.1.1.8, from the equine serum) was tested using the 5, 5'-dithiobis-(2-nitrobenzoic acid) (DTNB) method. DTNB would generate 5-mercapto-2-nitrobenzoic acid——a detectable chromophore at the 405~412 nm range. To evaluate the biological activity, the tested compounds were dissolved in dimethyl sulfoxide (DMSO) to an initial concentration of 0.1 mol/L, and then they were diluted into six different concentrations by a mixture of DMSO and EtOH (V:V=1:9), which could be tested to obtain the range of 20% to 80% enzyme inhibition for AChE or BuChE. For this purpose, in a 96 well plate, 26 μL of phosphate buffer (0.1 mol/L, pH=8.0), 30 μL of DTNB (0.01 mol/L), 4 μL of enzyme [5 U/mL of AChE or BuChE] and 10 μL of inhibitor solution or DMSO:EtOH mixture (V:V=1:9) was added. Then, 30 μL of substrate (0.01 mol/L, acetylthiocholine iodide or butyrylthiocholine iodide) was added to each well and the change of absorbance was measured at 410 nm for 2 min. Each experiment was done in triplicate. The IC50 values were determined graphically from inhibition curves (log inhibitor concentration vs. percent of inhibition).
4.3.2 AChE kinetic study with compound 3n
The kinetic studies were performed with compound 3n using the spectrometric Ellman’s method at different concentrations of the substrate acetylthiocholine iodide (0.111~1.111 mmol/L for each concentration of compound 3n). Four concentrations of 3n were used for the studies: 0, 1, 5 and 10 μmol/L. Each experiment was performed in triplicate. Vmax and Km values of the Michaelis-Menten kinetics were calculated by nonlinear regression from substrate-velocity curves. Lineweaver-Burk plots were calculated using linear regression.
4.4 Melting point measurement of compounds 3
The compounds 3 should be dried and ground. Take an appropriate amount of the sample and place it on a glass slide to make it distribution thin and even. Cover it with coverslip and lightly compact it, then place it in the center of the hot stage to observe the melting process and record the initial and complete melting temperature. The measurement is repeated three times to calculate the average value.
4.5 Modeling study
Compound 3n and donepezil with the acetylcholinesterase protein (PDB: 6O4W) and butyrylcholinesterase protein (PDB: 5K5E) were used as a model in the docking experiments. AutoDock 4.2 and UCSF Chimera were chosen to study the binding modes of the compound 3n, donepezil and their differences.
Supporting Information The 1H NMR and 13C NMR spectra of the target compounds. ClogP values of pyrano[2, 3-b]naphthoquinone derivatives 3. The supporting information is available free of charge via the Internet http://sioc-journal.cn/.
-
-
[1]
Bondi, M. W.; Edmonds, E. C.; Salmon, D. P. J. Int. Neuropsychol. Soc. 2017, 23, 818. doi: 10.1017/S135561771700100X
-
[2]
Cuetos, F.; Herrera, E.; Ellis, A. W. Neuropsychologia 2010, 48, 3329. doi: 10.1016/j.neuropsychologia.2010.07.017
-
[3]
Alzheimer's Association Alzheimers Dement. 2019, 15, 321.
-
[4]
Wimo, A.; Guerchet, M.; Ali, G. C.; Wu, Y. T.; Prina, A. M.; Winblad, B.; Jönsson, L.; Liu, Z.; Prince, M. Alzheimers Dement. 2016, 13, 1.
-
[5]
Scarpini, E.; Scheltens, P.; Feldman, H. Lancet Neurol. 2003, 2, 539. doi: 10.1016/S1474-4422(03)00502-7
-
[6]
Fan, L.; Mao, C.; Hu, X.; Zhang, S.; Yang, Z.; Hu, Z.; Sun, H.; Fan, Y.; Dong, Y.; Yang, J.; Shi, C.; Xu, Y. Front Neurol 2020, 10, 1312. doi: 10.3389/fneur.2019.01312
-
[7]
Contestabile, A. Behav. Brain Res. 2011, 221, 334. doi: 10.1016/j.bbr.2009.12.044
-
[8]
Enz, A.; Amstutz, R.; Boddeke, H.; Gmelin, G.; Malanowski, J. Prog. Brain Res. 1993, 98, 431. doi: 10.1016/S0079-6123(08)62429-2
-
[9]
Davies, P.; Maloney, A. J. Lancet 1976, 2, 1403.
-
[10]
Babu, M. A.; Lakshmi, M.; Vasanthanathan, P. G.; Kaskhedikar, S. Indian J. Pharm. Sci. 2005, 67, 1.
-
[11]
Peauger, L.; Azzouz, R.; Gembus, V.; Ţînţaş, M. L.; Sopková-de Oliveira Santos, J.; Bohn, P.; Papamicaël, C.; Levacher, V. J. Med. Chem. 2017, 60, 5909. https://pubmed.ncbi.nlm.nih.gov/28613859/
-
[12]
(a) Auld, D. S.; Kornecook, T. J.; Bastianetto, S.; Quirion, R. Prog. Neurobiol. 2002, 68, 209.
(b) Mohammad, D.; Chan, P.; Bradley, J.; Lanctôt, K.; Herrmann, N. Expert Opin. Drug Saf. 2017, 16, 1009. -
[13]
(a) Smith, D. A. Am. J. Health-Syst. Pharm. 2009, 66, 899.
(b) Herrmann, N.; Chau, S. A.; Kircanski, I.; Lanctôt, K. L. Drugs 2011, 71, 2031.
(c) Misra, S.; Medhi, B. Neurol. Sci. 2013, 34, 831. -
[14]
Brewster, J. T.; Dell'Acqua, S.; Thach, D. Q.; Sessler, J. L. ACS Chem. Neurosci. 2019, 10, 155. doi: 10.1021/acschemneuro.8b00517
-
[15]
(a) Luo, Z.; Sheng, J.; Sun, Y.; Lu, C.; Yan, J.; Liu, A.; Luo, H. B.; Huang, L.; Li, X. J. Med. Chem. 2013, 56, 9089.
(b) Graham, W. V.; Bonito-Oliva, A.; Sakmar, T. P. Annu. Rev. Med. 2017, 68, 413. -
[16]
Kuhl, D. E.; Koeppe, R. A.; Snyder, S. E.; Minoshima, S.; Frey, K. A.; Kilbourn, M. R. Ann. Neurol. 2006, 59, 13. doi: 10.1002/ana.20672
-
[17]
Akiko, K. J.; Todd, E.; Keith, D. G.; Abdelrahman, S. M.; Mi, H. L.; Sylvie, G. T. Chem. Sci. 2013, 4, 4137. doi: 10.1039/c3sc51902c
-
[18]
Li, S. Y.; Jiang, N.; Xie, S. S.; Wang, K. D.; Wang, X. B.; Kong, L. Y. Org. Biomol. Chem. 2014, 12, 801. doi: 10.1039/C3OB42010H
-
[19]
Xie, S. S.; Wang, X.; Jiang, N.; Yu, W.; Wang, K. D.; Lan, J. S.; Li, Z. R.; Kong, L. Y. Eur. J. Med. Chem. 2015, 95, 153. doi: 10.1016/j.ejmech.2015.03.040
-
[20]
Demir Özkay, Ü.; Can, Ö. D.; Sağlık, B. N.; Acar Çevik, U.; Levent, S.; Özkay, Y.; Ilgın, S.; Atlı, Ö. Bioorg. Med. Chem. Lett. 2016, 26, 5387. doi: 10.1016/j.bmcl.2016.10.041
-
[21]
Azzouz, R.; Peauger, L.; Gembus, V.; Ţînţaş, M. L.; Sopková-de Oliveira Santos, J.; Papamicaël, C.; Levacher, V. Eur. J. Med. Chem. 2018, 145, 165. doi: 10.1016/j.ejmech.2017.12.084
-
[22]
Singh, M.; Silakari, O. RSC Adv. 2016, 6, 108411. doi: 10.1039/C6RA17678J
-
[23]
Zheng, J.; He, M.; Xie, B.; Yang, L.; Hu, Z.; Zhou, H. B.; Dong, C. Org. Biomol. Chem. 2018, 16, 472. doi: 10.1039/C7OB02794J
-
[24]
Martín-Acosta, P.; Haider, S.; Amesty, Á.; Aichele, D.; Jose, J.; Estévez-Braun, A. Eur. J. Med. Chem. 2018, 144, 410. doi: 10.1016/j.ejmech.2017.12.058
-
[25]
Wang, X. H.; Zhang, X. H.; Tu, S. J.; Shi, F.; Zou, X.; Yan, S.; Han, Z. G.; Hao, W. J.; Cao, X. D.; Wua, S. S. J. Heterocycl. Chem. 2009, 46, 832. doi: 10.1002/jhet.153
-
[26]
Khan, N.; Pal, S.; Karamthulla, S.; Choudhury, L. H. RSC Adv. 2015, 45, 3732. https://www.researchgate.net/publication/258374792_ChemInform_Abstract_Imidazole_as_Organocatalyst_for_Multicomponent_Reactions_Diversity_Oriented_Synthesis_of_Functionalized_Hetero-_and_Carbocycles_Using_in_situ-Generated_Benzylidenemalononitrile_Der
-
[27]
Ellman, G. L.; Courtney, K. D.; Andres, V. Jr; Feather-Stone, R. M. Biochem. Pharmacol. 1961, 7, 88. doi: 10.1016/0006-2952(61)90145-9
-
[28]
Maleki, B.; Babaee, S.; Tayebee, R. Appl. Organomet. Chem. 2015, 29, 408. doi: 10.1002/aoc.3306
-
[29]
Sameem, B.; Saeedi, M.; Mahdavi, M.; Nadri, H.; Moghadam, F. H.; Edraki, N.; Khan, M. I.; Amini, M. Bioorg. Med. Chem. 2017, 25, 3980. https://xueshu.baidu.com/usercenter/paper/show?paperid=1c72bce0462ab364e4008c43db19548d&site=xueshu_se
-
[30]
Czarnecka, K.; Chufarova, N.; Halczuk, K.; Maciejewska, K.; Girek, M.; Skibiński, R.; Jończyk, J.; Bajda, M.; Kabziński, J.; Majsterek, I.; Szymański, P. Eur. J. Med. Chem. 2018, 145, 760. doi: 10.1016/j.ejmech.2018.01.014
-
[31]
Dgachi, Y.; Sokolov, O.; Luzet, V.; Godyń, J.; Panek, D.; Bonet, A.; Martin, H.; Iriepa, I.; Moraleda, I.; García-Iriepa, C.; Janockova, J.; Richert, L.; Soukup, O.; Malawska, B.; Chabchoub, F.; Marco- Contelles, J.; Ismaili, L. Eur. J. Med. Chem. 2017, 126, 576. doi: 10.1016/j.ejmech.2016.11.050
-
[32]
Eghtedari, M.; Sarrafi, Y.; Nadri, H.; Mahdavi, M.; Moradi, A.; Homayouni Moghadam, F.; Emami, S.; Firoozpour, L.; Asadipour, A.; Sabzevari, O.; Foroumadi, A. Eur. J. Med. Chem. 2017, 128, 237. https://www.sciencedirect.com/science/article/abs/pii/S0223523417300600
-
[33]
Kavita, J.; Saikat, C.; Kuntal, P.; Kalpataru, D. New J. Chem. 2019, 43, 1299. doi: 10.1039/C8NJ04219E
-
[34]
Aniruddha, D.; Nagaraj, A.; Amarajothi, D.; Shyam, B. Microporous Mesoporous Mater. 2019, 284, 459. doi: 10.1016/j.micromeso.2019.04.057
-
[35]
Li, C. X.; Zhong, D. D.; Huang, X. Q.; Shen, G. D.; Li, Q.; Du, J. Y.; Li, Q. L.; Wang, S. N.; Li, J. K.; Dou, J. M. New J. Chem. 2019, 43, 5813. doi: 10.1039/C8NJ06460A
-
[36]
Guo, F.; Su, C. H.; Chu, Z. P.; Zhao, M. H. J. Solid State Chem. 2019, 277, 25. doi: 10.1016/j.jssc.2019.05.036
-
[1]
-
表 1 AChE inhibition data, IC50 against AChE and BuChE of pyrano[2, 3-b]naphthoquinone derivatives 3
Entry Compd. AChE inhibition/% IC50c/(μmol•L-1) 20 μmol/La 50 μmol/Lb AChE BuChE 1 3a 59.66 67.87 10.40±0.90 >200 2 3b 55.94 67.99 6.09±0.50 >200 3 3c 60.25 69.12 2.37±0.30 >200 4 3d 51.14 73.52 20.70±1.80 >200 5 3e 63.90 74.83 4.31±0.10 >200 6 3f 80.80 87.59 3.67±0.10 >200 7 3g 67.79 86.52 6.03±0.10 >200 8 3h 68.46 89.61 6.50±0.08 >200 9 3i 68.80 82.38 12.70±0.40 >200 10 3j 32.35 62.70 37.10±2.50 >200 11 3k 80.32 86.54 3.82±0.20 >200 12 3l 67.22 81.77 11.80±1.27 >200 13 3m 66.68 80.63 4.17±0.59 >200 14 3n 70.90 74.91 1.22±0.12 >200 15 3o 57.17 64.96 2.25±0.06 >200 16 3p 63.83 71.41 5.29±0.02 >200 17 3q 80.15 78.78 4.36±1.09 >200 18 3r 22.70 27.54 >50 >200 19 3s 61.64 67.57 4.43±0.91 >200 20 3t 59.63 63.27 19.40±0.42 >200 21 3u 35.89 65.47 22.10±1.98 >200 22 3v 36.33 43.51 >50 >200 23 3w 35.89 65.47 25.50±2.26 >200 24 3x 20.99 34.47 >100 >200 25 Donepezil — — 0.03±0.001 29.7±2.46 a Data are represented as AChE inhibition (%) at 20 μmol/L. b Data are represented as AChE inhibition (%) at 50 μmol/L. c Data are represented as mean±SD. All experiments were independently carried out at least three times. -

计量
- PDF下载量: 9
- 文章访问数: 1206
- HTML全文浏览量: 135