欧芹酚甲醚肟酯类化合物的制备及其抗乙酰胆碱酯酶活性研究

为研发具有AChE双位点抑制的新型乙酰胆碱酯酶抑制剂,本论文以香豆素类化合物欧芹酚甲醚为先导化合物,通过氧化、还原、酯化等反应设计合成了14个欧芹酚甲醚肟酯衍生物,并经核磁共振波谱(NMR)和质谱(MS)进行结构确证。采用Ellman法测定了目标化合物在1μmol·mL^-1下对乙酰胆碱酯酶的抑制活性,结果显示目标化合物4i和4n对乙酰胆碱酯酶的抑制率分别达到61.6%和51.2%,且与化合物浓度呈正相关。分子对接分析表明化合物4i可以和AChE的催化位点和外周阴离子位点结合。     

胆碱酯酶抑制剂萘酰亚胺衍生物的合成与聚集诱导发光性质

阿尔茨海默症(Alzheimer’s disease, AD)是一种神经退行性疾病,严重影响老年人的生活质量,目前治疗AD的药物主要是胆碱酯酶抑制剂,如多奈哌齐、卡巴拉汀等.本文基于多奈哌齐结构,设计合成了一系列新的萘酰亚胺衍生物并进行了活性评价.结果表明,所合成的化合物均对乙酰胆碱酯酶(AChE)有选择性抑制,其中2-((1-(3-甲氧基苄基)哌啶-4-基)甲基)-1H-苯并异喹啉-1,3(2H)-二酮(4k)的抑制活性最强,IC₅₀值为4.43μmol·L^-1,优于对照药物卡巴拉汀.酶动力学及分子对接表明4k能够同时作用于ACh E的催化活性位点和外周结合位点,并且4k对SH-SY5Y和PC12细胞毒性较低.此外,这些化合物均显示出典型的聚集诱导发光(AIE)性质,可能与萘酰亚胺分子内旋转受阻机制有关.     

4-N-苯胺基喹啉衍生物的合成及胆碱酯酶抑制活性

设计合成了一系列4-N-苯胺基喹啉类衍生物,采用Ellman法测定了目标化合物对乙酰胆碱酯酶(AChE)和丁酰胆碱酯酶(BChE)的抑制活性.结果表明,当喹啉环上连有伸长的吡啶季铵盐片段时,可显著提高目标化合物的胆碱酯酶抑制作用.化合物16对AChE和BChE具有明显的双重抑制作用,其IC_(50)值分别为0.92和14.20μmol/L,抑制效果强于阳性对照药加兰他敏.     

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（英文）

设计、合成了一系列新型吡喃并[2,3-b]萘醌类衍生物.抗胆碱抑制活性测试显示,与丁酰胆碱酯酶(BuChE)相比,大部分化合物显示出对乙酰胆碱酯酶(AChE)的高选择性和良好的抑制活性.其中活性最好的化合物(2-氨基-4-(3-氰基苯基)-5,10-二氧代-5,10-二氢-4H-苯并[g]亚甲基-3-甲腈)(3n),其AChE抑制活性IC_(50)值为1.22μmol/L,比BuChE高164倍.此外,分子对接模拟研究为理解这些化合物的作用机制、效力及选择性提供了理论支持.这些新型有效且高度选择性的AChE抑制剂的发现为开发阿尔茨海默症的潜在治疗药物提供了先导化合物.     

基于固定化酶的乙酰胆碱酯酶抑制剂体外筛选模型的建立

以可重复使用的固定化酶代替游离态酶, 建立一种基于比色分析的乙酰胆碱酯酶(AChE)抑制剂体外筛选新模型. 采用以氨基化硅胶为载体固定的AChE优化了实验条件, 用AChE抑制剂阳性对照物他克林和毒扁豆碱对该模型进行验证, 还对模型技术参数进行评价, 并将新模型用于单体化合物及天然产物粗提物AChE抑制活性评价. 结果表明, 最佳实验条件为: 固定化酶用量55 μL, 底物浓度5 mmol/L, 甲醇、 乙醇及体积分数不高于6%的二甲基亚砜水溶液均可作为样品溶剂; 模型验证及模型技术参数评价结果良好, 该模型对AChE抑制剂筛选有较好的特异性和灵敏度, 可用于筛选AChE抑制剂. 该模型具有适用性强、 固定化酶可重复使用及结果可靠等优点, 是单体化合物及天然产物粗提物AChE抑制剂活性评价的有效方法.     

欧芹酚甲醚腙类衍生物的合成及抗乙酰胆碱酯酶活性研究

以香豆素类化合物欧芹酚甲醚为先导化合物,设计合成了15个欧芹酚甲醚腙类衍生物,所有目标化合物经熔点、¹H NMR和MS进行结构确证。体外抑制乙酰胆碱酯酶活性结果表明,在100μmol/L浓度下,目标化合物3d对乙酰胆碱酯酶具有较强的抑制活性,其抑制率达到66.1%。初步构效关系表明,在欧芹酚甲醚腙类化合物的苯环上引入斥电子基能提高对乙酰胆碱酯酶的抑制活性。分子对接表明化合物3d可以和乙酰胆碱酯酶的催化活性中心部位显著结合。     

基于AChE和BuChE靶标的查尔酮衍生物的合成及生物活性研究

为了探寻治疗阿尔茨海默症（AD）的潜在双靶标活性化合物,本研究设计合成了5个新的查尔酮衍生物,并对它们的生物活性进行了评价。酶活性测定结果表明,化合物C1～C5对乙酰胆碱酯酶（AChE）和丁酰胆碱酯酶（BuChE）均具有显著的抑制活性。其中,化合物C4对AChE和BuChE有明显的抑制作用,IC₅₀分别为6.00和2.63μmol/L。细胞实验表明,化合物C4对Glu、H₂O₂和Aβ_1-42诱导的神经母细胞瘤SH-SY5Y损伤具有显著的保护作用,且呈剂量依赖性。Y迷宫和Morris水迷宫研究表明,C4能改善东莨菪碱诱导的小鼠的认知障碍和背景记忆能力,且对小鼠自主活动无影响,具有毒副作用小的优点。此外,C4能显著降低小鼠皮层总胆碱酯酶和丙二醛含量,提高谷胱甘肽过氧化物酶和超氧化物歧化酶含量。分子模拟显示,化合物C4能够结合AChE和BuChE的催化活性位点和外周阴离子位点。因此,化合物C4可能是一种很有前途的双靶向活性分子,值得做进一步的结构优化和活性评价,以期发现更安全高效的AD治疗药物。     

新型20(S)-O-取代苯甲酸-7-苯甲酰基喜树碱酯的合成及其抗肿瘤活性

以20(S)-喜树碱为起始原料,对其进行结构修饰,在7-位导入苯甲酰基,在20-位羟基上导入取代苯甲酰基,设计并合成了11个新的20(S）-O-取代苯甲酸-7-苯甲酰基喜树碱酯化合物（4a~4k）,其结构经¹H NMR, IR,MS(ESI)和元素分析表征。采用MTT法初步考察了目标化合物4a～4j对人胃癌细胞（BGC-823）、人乳腺癌细胞（MDA-MB-231）、人肺腺癌细胞（H460）及人肝癌细胞（Bel-7404）的体外抑制活性。结果表明：化合物4a、 4g和4h具有一定的体外抑瘤活性。在药物浓度为10 μmol·L^-1时,4a对MDA-MB-231细胞和H460细胞的抑制率分别为50.42%和54.40%, 4g〗对MDA-MB-231细胞的抑制率为69.91%, 4h对H460细胞抑制率为52.34%。     

基于他克林的乙酰胆碱酯酶抑制剂的3D-QSAR研究及虚拟筛选

本文通过对58个他克林派生物乙酰胆碱酯酶抑制剂分子进行建模分析,研究其结构与活性的关系,并通过虚拟筛选方法获得一系列潜在AChE抑制剂双位点分子。首先将一系列他克林二联体化合物与AChE晶体结构对接,获得化合物的活性构象,以此进行建模分析,建立结构与活性之间的三维定量构效关系。所得模型CoMFA、CoMSIA、TopomerCoMFA的交叉验证系数分别为0.510、0.702、0.571,非交叉验证系数为0.998、0.988、0.794,测试集r_(pred)~2为0.750、0.742、0.766,所得模型具有良好的预测性,由此可以为设计高活性的新分子提供理论基础。然后,使用Topomer search对ZINC数据库中的125909分子进行虚拟筛选,得到891个具有潜在AChE抑制活性的分子。最后,对这891个分子进行分子对接,观察分子与晶体结构的结合情况,筛选得到66个具有高选择性的双位点AChE抑制剂分子。     

1-芳基-3-(3-(4-氧香豆素基)苯基)硫脲衍生物的合成及抑菌活性

为了筛选出具有较高抑菌活性的含香豆素的硫脲类衍生物,本文以4-羟基香豆素为原料,经氯化、醚化、异硫氰酸化和加成反应合成了一系列1-芳基-3-(3-(4-氧香豆素基)苯基)硫脲衍生物,其结构经红外光谱(IR)、核磁共振谱(NMR)和质谱(MS)等技术手段进行了表征。结果表明,目标化合物对水稻白叶枯菌和柑橘溃疡菌均具有较好的抑制活性。其中化合物4k、4l、4m和4n抑制水稻白叶枯菌活性EC_(50)值分别为137.42、131.05、129.23和117.43 mg/L,优于对照药剂噻菌铜的活性(195.24 mg/L);化合物4k、4l、4m和4n抑制柑橘溃疡菌活性EC_(50)值分别为97.02、94.31、102.28和90.52 mg/L,优于噻菌铜的活性(120.25 mg/L)。     

重组嗜热β-葡萄糖苷酶转化稀有人参皂苷Rd和CK

利用高效液相色谱(HPLC)法, 对重组嗜热β-葡萄糖苷酶(Fpglu1)转化稀有人参皂苷(Rd和CK)进行研究, 并表征了其催化动力学参数. 利用同源模建和分子动力学模拟等生物信息学技术, 探究了Fpglu1转化人参皂苷的结构基础及其相互作用. 结果表明, Fpglu1能够水解人参总皂苷生成稀有皂苷Rd和CK, 其催化人参皂苷Rb₁, Rb₂和Rc的K_m值分别为0.318, 1.840和5.269 mmol/L; 酶的转换数(k_cat)值分别为144.191, 0.572和0.011 s^-1. 当转化时间分别为6和102 h时, Rd和CK的产率达到最大, 分别为60%和93%. 通过对该酶的结构预测及皂苷分子的对接研究发现, 底物位于由疏水性氨基酸构成的底物口袋中, 氨基酸残基Glu194和Glu367是参与催化作用的关键, 且实验测得的酶促反应动力学参数(K_m)与对接的相互作用能量值存在线性关系.     

Synthesis and biological evaluation of novel 1, 2, 3-benzotriazin-4-one derivatives as leukotriene A4 hydrolase aminopeptidase inhibitors

A series of novel 1,2,3-benzotriazin-4-one derivatives were designed,synthesized and their inhibitory activities against leulcotriene A_4 hydrolase aminopeptidase in vitro were evaluated.Many compounds showed moderate to good activities at the concentration of 10 μmol/L.Among them,compound Ⅳ-16 exhibited the highest inhibitory activity up to 80.6% with an IC_(50) of 1.30 ± 0.20 μmol/L The compound Ⅳ-16 was also tested the proliferation inhibitory activities in THP1 human AML cell line and its binding model with LTA_4H enzyme by molecular docking was studied.It indicated that 1,2,3-benzotriazin-4-one was a promising scaffold for further study.The relationship between structure and inhibitory activity was also preliminarily discussed.     

7-二乙氨基香豆素腙及酰腙衍生物的制备及乙酰胆碱酯酶抑制活性研究

本论文采用亚活性结构拼接的方法,以7-N,N-二乙氨基-4-甲基香豆素为原料,经二氧化硒氧化,然后与取代肼和取代酰肼反应,合成了13个7-N,N-二乙氨基香豆素腙及酰腙衍生物。所有目标化合物经1H NMR和MS进行结构确证。体外抑制乙酰胆碱酯酶活性结果表明,目标化合物4a和4c对乙酰胆碱酯酶具有较强的抑制活性,其IC50值分别为42. 89和90. 32μmol/L。初步构效关系研究表明,酰腙衍生物对乙酰胆碱酯酶的抑制活性比腙类衍生物好。     

Design and synthesis of novel triazole derivatives containing γ-lactam as potential antifungal agents

A series of novel triazole derivatives containing γ-lactam were designed and synthesized, and their structures were confirmed by ~1H NMR,~(13)CNMR and HRMS. The in vitro antifungal activities of the target compounds were evaluated. The results showed that all of the compounds exhibited stronger activity against the six clinically important fungi tested than fluconazole. 3D and 3E showed comparative activity against the fungi tested except for Candida glabrata and Aspergillus fumigatus as voriconazole. In addition,the docking model for 2A and CYP51 was investigated.     

1,4-戊二烯-3-酮肟醚类化合物的合成及生物活性

将1,4-戊二烯-3-酮结构中的羰基转变为肟醚基团,合成了7个1,4-戊二烯-3-酮肟醚类化合物,其结构经1H NMR、13C NMR、IR和ESI-MS表征。生物活性测试结果表明,在药剂浓度为500mg/L时,化合物4c、4f和4g对烟草花叶病毒(TMV)在治疗作用方面的抑制率为49.6%、53.6%和47.4%,与病毒唑(45.2%)相当;化合物4b、4c、4f和4g对TMV在保护作用方面的抑制率分别为57.2%、58.4%、58.9%和59.0%,稍低于病毒唑(61.8%);化合物4a、4b、4c和4g对TMV在钝化作用方面的抑制率分别为95.5%、92.6%、95.0%和89.5%,优于对照药病毒唑(87.9%)。这些结果表明,1,4-戊二烯-3-酮肟醚类化合物对植物病毒表现有良好抑制作用,在其结构基础上进行适当的优化,有望得到具有良好抗植物病毒活性的先导化合物。     

Structural Insights and Docking Analysis of Adamantane-Linked 1,2,4-Triazole Derivatives as Potential 11β-HSD1 Inhibitors

The solid-state structural analysis and docking studies of three adamantane-linked 1,2,4-triazole derivatives are presented. Crystal structure analyses revealed that compound 2 crystallizes in the triclinic P-1 space group, while compounds 1 and 3 crystallize in the same monoclinic P2₁/c space group. Since the only difference between them is the para substitution on the aryl group, the electronic nature of these NO₂ and halogen groups seems to have no influence over the formation of the solid. However, a probable correlation with the size of the groups is not discarded due to the similar intermolecular disposition between the NO₂/Cl substituted molecules. Despite the similarities, CE-B3LYP energy model calculations show that pairwise interaction energies vary between them, and therefore the total packing energy is affected. HOMO-LUMO calculated energies show that the NO₂ group influences the reactivity properties characterizing the molecule as soft and with the best disposition to accept electrons. Further, in silico studies predicted that the compounds might be able to inhibit the 11β-HSD1 enzyme, which is implicated in obesity and diabetes. Self- and cross-docking experiments revealed that a number of non-native 11β-HSD1 inhibitors were able to accurately dock within the 11β-HSD1 X-ray structure 4C7J. The molecular docking of the adamantane-linked 1,2,4-triazoles have similar predicted binding affinity scores compared to the 4C7J native ligand 4YQ. However, they were unable to form interactions with key active site residues. Based on these docking results, a series of potentially improved compounds were designed using computer aided drug design tools. The docking results of the new compounds showed similar predicted 11β-HSD1 binding affinity scores as well as interactions to a known potent 11β-HSD1 inhibitor.     

Modeling ligand-receptor interaction for some MHC class II HLA-DR4 peptide mimetic inhibitors using several molecular docking and 3D QSAR techniques

The ligand-receptor interaction between some peptidomimetic inhibitors and a class II MHC peptide presenting molecule, the HLA-DR4 receptor, was modeled using some three-dimensional (3D) quantitative structure-activity relationship (QSAR) methods such as the Comparative Molecular Field Analysis (CoMFA), Comparative Molecular Similarity Indices Analysis (CoMSIA), and a pharmacophore building method, the Catalyst program. The structures of these peptidomimetic inhibitors were generated theoretically, and the conformations used in the 3D QSAR studies were defined by docking them into the known structure of HLA-DR4 receptor through the GOLD, GLIDE Rigidly, GLIDE Flexible, and Xscore programs. Some of the parameters used in these docking programs were selected by docking an X-ray ligand into the receptor and comparing the root-means-square difference (RMSD) computed between the coordinates of the X-ray and docked structure. However, the goodness of a docking result for docking a series of peptidomimetic inhibitors into the HLA-DR4 receptor was judged by comparing the Spearman's rank correlation coefficient computed between each docking result and the activity data taken from the literature. The best CoMFA and CoMSIA models were constructed using the aligned structures of the best docking result. The CoMSIA was conducted in a stepwise manner to identify some important molecular features that were further employed in a pharmacophore building process by the Catalyst program. It was found that most inhibitors of the training set were accurately predicted by the best pharmacophore model, the Hypo1 hypothesis constructed. The deviation or conflict found between the actual and predicted activities of some inhibitors of both the training and the test sets were also investigated by mapping the Hypo1 hypothesis onto the corresponding structures of the inhibitors.     

新型桃金娘烯醛基双酰胺-噻二唑化合物的合成及其抗真菌活性

以α-蒎烯为起始原料,经氧化制得桃金娘烯酸（3）;脂肪族二酸在POCl₃作用下与氨基硫脲经脱水环合制得脂肪族双噻二唑（4a~4h）; 4a~4h分别与3经脱水反应合成了8个新型的桃金娘烯醛基双酰胺噻二唑化合物（5a~5h）,其结构经¹H NMR, ¹³C NMR, IR和ESI-MS表征。抗真菌活性测试结果表明,在用药量为50 μg·mL^-1时,5a~5h对黄瓜枯萎病菌、花生褐斑病菌、苹果轮纹病菌、番茄早疫病菌和小麦赤霉病菌均有一定的抑制作用,其中桃金娘烯醛-辛二酸基双酰胺-噻二唑(5f)对苹果轮纹病菌的抑制率为60.3%,桃金娘烯醛-丁二酸基双酰胺-噻二唑(5b)和桃金娘烯醛-癸二酸基双酰胺-噻二唑(5h)对小麦赤霉病菌的抑制率分别为52.8%和54.4%。     

GC-MS,LC-MS/MS,Docking and Molecular Dynamics Approaches to Identify Potential SARS-CoV-2 3-Chymotrypsin-Like Protease Inhibitors from Zingiber officinale Roscoe

This study aims to identify and isolate the secondary metabolites of Zingiber officinale using GC-MS, preparative TLC, and LC-MS/MS methods, to evaluate the inhibitory potency on SARS-CoV-2 3 chymotrypsin-like protease enzyme, as well as to study the molecular interaction and stability by using docking and molecular dynamics simulations. GC-MS analysis suggested for the isolation of terpenoids compounds as major compounds on methanol extract of pseudostems and rhizomes. Isolation and LC-MS/MS analysis identified 5-hydro-7, 8, 2′-trimethoxyflavanone (9), (E)-hexadecyl-ferulate (1), isocyperol (2), N-isobutyl-(2E,4E)-octadecadienamide (3), and nootkatone (4) from the rhizome extract, as well as from the leaves extract with the absence of 9. Three known steroid compounds, i.e., spinasterone (7), spinasterol (8), and 24-methylcholesta-7-en-3β-on (6), were further identified from the pseudostem extract. Molecular docking showed that steroids compounds 7, 8, and 6 have lower predictive binding energies (MMGBSA) than other metabolites with binding energy of −87.91, −78.11, and −68.80 kcal/mole, respectively. Further characterization on the single isolated compound by NMR showed that 6 was identified and possessed 75% inhibitory activity on SARS-CoV-2 3CL protease enzyme that was slightly different with the positive control GC376 (77%). MD simulations showed the complex stability with compound 6 during 100 ns simulation time.     

Synthesis,In vitro Coagulation Activities and Molecular Docking Studies on Three L-Histidine Amide Derivatives

Three novel L-histidine amide derivatives were synthesized and the corresponding chemical structures were characterized by means of melting point analysis, IR, MS, ¹H NMR as well as ¹³C NMR. The coagulation acti- vities of the compounds were evaluated by an MOE(molecular operating environment) docking technique and coagulation test. The results obtained from molecular docking show that the interactions between the compounds and thrombin exhibit procoagulant activity in combination with an improved combinatory effect. Moreover, the results of in vitro coagulation tests show that the L-histidine amide derivatives feature coagulant activities in common coagulation pathways. Compared with the blank control group, the optimal shortening rates of compounds 1―3 were 39.08%(0.5 mmol/L), 22.94%(1.0 mmol/L) and 15.38%(0.0625 mmol/L), respectively.