一种新型化合物的合成及对PC12高分化细胞的神经保护作用（英文）

研究了一种新合成化合物(3h8b)的神经保护作用及初步分子机制.该化合物具有缓解神经毒素6-羟基多巴胺(6-OHDA)和L-谷氨酸(L-Glu)诱导高分化PC12细胞损伤的作用.可有效改善神经毒素对高分化PC12细胞的损伤,明显增强细胞活力,降低细胞核凋亡比率,抑制细胞内钙离子过载.经3h8b处理之后,神经毒素引起的细胞内线粒体膜电位异常显著恢复.进一步的实验表明,3h8b可以逆转神经毒素造成的Bcl-2和Bcl-x L抑制性表达.实验证实3h8b通过线粒体相关途经对高分化PC12细胞起保护作用,为通过化学合成法获得具有治疗神经退行性病变作用的新型化合物提供了理论依据.     

硫（醇）代N-对甲苯基/苯基磺酰基氨基羧酸酯类化合物的合成与神经营养活性

根据官能团的组合,设计合成了硫（醇）代N-对甲苯基/苯基磺酰基氨基羧酸酯类化合物,这些化合物的结构¹H NMR, ¹³C NMR, MS 和 HRMS证实,初步的实验结果表明：在10μg/mL浓度下,化合物4a, 4b, 4d, 5c, 5d, 5g, 6b和 6d显示对PC12细胞缺氧损伤具有显著的保护作用,化合物4c, 5b和 6c显示对PC12细胞缺氧损伤具有一定的保护作用;在5μg/mL浓度下,4d和 6d显示对PC12细胞缺氧损伤具有一定的保护作用;初步的实验也表明：在10μg/mL浓度下,4c, 5a, 5c, 5d, 5e和 6b显示对PC12细胞具有一定的促分化作用。     

熊果酸半乳糖苷偶联物的合成及保肝活性

通过改进的Koenigs-knorr法在熊果酸3位和28位进行半乳糖苷化得到6个化合物. 通过MTT法考察了上述化合物对大鼠肝干细胞样上皮细胞WB-F344 的作用, 发现化合物12b和12e可明显提高WB-F344细胞的成活率. 体内实验结果表明, 化合物12b, 12d和12f在刀豆蛋白引起的小鼠急性免疫性肝损伤模型上, 除化合物12d对小鼠血清谷草转氨酶升高具有一定程度的降低作用外, 其余化合物均未见对谷草和谷丙转氨酶的升高表现出明显的保护作用.     

脑源性神经营养因子保护GdCl_3对神经元与星形胶质细胞共培养体系中的毒性作用

星形胶质细胞和神经元之间的相互作用对于神经元的生长及调节过程十分重要.脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)是脑内广泛分布的一类神经营养因子,除神经元分泌外,星形胶质细胞分泌BDNF功能在神经细胞损伤和神经退行性病变过程起重要作用.本实验之前的研究表明,Gd引起的神经元死亡伴随线粒体功能的损伤,如线粒体呼吸链功能障碍,ATP的合成量的减少和线粒体膜电位的降低.同时细胞内ROS水平的升高.抑制细胞氧化应激水平能够降低Gd暴露后神经元的死亡.神经元产生BDNF功能的影响可能和ROS介导的细胞毒性有密切的关系.因此,Gd对神经元分泌BDNF功能的影响可能是其导致神经毒性的又一因素,本实验研究了Gd对单独培养的神经元以及神经元与星形胶质细胞共培养体系的作用.通过测定细胞上清乳酸脱氢酶(LDH)的活性和观察PI阳性染色的细胞数目来分析细胞死亡率.结果表明,单独培养的神经元暴露GdCl3 12h后,细胞上清LDH的活力显著增加(约40%).但是,与星形胶质细胞共培养或者单独培养的星形胶质细胞暴露于GdCl3后并没有LDH的显著增加.BDNF能够降低Gd导致的细胞上清LDH的升高.同时,单独培养的神经元暴露于GdCl3后,PI阳性染色的细胞数目明显增多.与星形胶质细胞共培养或者BDNF的干预均能够降低Gd导致的PI阳性染色数目的增加.因此,Gd引起的神经元死亡和其对细胞产生BDNF功能的影响有关系,星形胶质细胞能够够减轻Gd对神经元的毒性作用.用DCFH-DA标记细胞内ROS,用激光共聚焦显微镜(CLSM)观察不同细胞用GdCl3孵育后细胞内ROS荧光强度.结果提示,用20μM的GdCl3孵育不同细胞12h后,单独培养的神经元细胞内ROS水平显著升高.但是与星形胶质细胞共培养或者BDNF处理的神经元,ROS水平并没有明显的升高.因此,与星形胶质细胞共培养或者BDNF的干预能够降低Gd引起的神经元的死亡.BDNF能够降低Gd诱导的神经元ROS水平的升高以及细胞的死亡.因此,对细胞产生BDNF功能的影响可能是Gd导致神经元细胞毒性的主要因素之一.实验结果显示,GdCl3孵育单独培养的神经元不同时间后,BDNF mRNA和蛋白表达随着GdCl3孵育时间的延长而降低.当GdCl3孵育时间达到12h时,表达量降低了约25%.当GdCl3孵育时间达到24h时,对BDNF表达的影响进一步加强.但与单独培养的神经元相比,当神经元与星形胶质共培养时,20μM的GdCl3孵育细胞并没有引起BDNF表达的明显降低.因此,星形胶质细胞能够通过调节BDNF的表达对神经元的损伤起到保护作用,神经元中BDNF水平的维持可能来源于直接星形胶质细胞分泌的BDNF,或者由外源性BDNF诱导了神经元中BDNF的表达.我们提出了一种可能的分子机制,将有助于理解钆化合物对神经细胞的毒理作用.     

高效液相色谱-电喷雾-串联质谱研究四溴双酚A双(2-羟乙基醚)诱导大鼠嗜铬细胞瘤细胞呼吸链氧化磷酸化和能量代谢紊乱

四溴双酚A衍生物的毒理学研究亟须开展。已有研究发现四溴双酚A双(2-羟乙基醚)(tetrabromobisphenol A bis(2-hydroxyethyl ether),TBBPA-BHEE)可诱导大鼠嗜铬细胞瘤细胞(PC12)活性氧(reactive oxygen species,ROS)的生成。然而TBBPA-BHEE对PC12细胞线粒体呼吸链氧化磷酸化过程的干扰机制尚不明确,TBBPA-BHEE是否通过破坏线粒体功能干扰细胞能量代谢亟待进一步探讨。建立了基于HPLC-ESI-MS/MS分析PC12细胞内ATP、ADP、AMP及cAMP(cyclic AMP)浓度的方法,在此基础上评价了TBBPA-BHEE暴露对PC12线粒体呼吸链氧化磷酸化过程及能量代谢的影响。研究发现,TBBPA-BHEE可加速PC12线粒体呼吸链氧化磷酸化过程;TBBPA-BHEE诱导的PC12线粒体功能紊乱可引起细胞能量代谢紊乱。一方面揭示了TBBPA-BHEE对PC12潜在的毒性作用机制,另一方面也证实HPLC-ESI-MS/MS是研究细胞线粒体呼吸链氧化磷酸化及能量代谢过程的有力工具。     

喹啉-多胺衍生物的合成及其活性评价

设计合成了13个喹啉-多胺衍生物,产物结构均经1H NMR,ESI-MS和元素分析确认.采用噻唑蓝(MTT)法测试了化合物对连二亚硫酸钠损伤肾上腺嗜铬细胞瘤(PC12)细胞的抑制作用,结果表明所有目标化合物对PC12细胞损伤均有较好的抑制作用,部分化合物优于尼莫地平,其中化合物T6和T7的活性最强,10μmol/L浓度下损伤抑制率分别为28.04%和27.58%.     

柚皮苷双席夫碱的合成及生物活性研究

将柚皮苷与1,3-丙二胺缩合得到一种柚皮苷双席夫碱。通过Th T实验、MTT实验、SOD检测以及细胞内ROS检测考察了该化合物生物活性。实验结果发现,该化合物能有效阻止Cu~(2+)诱导的Aβ_(1-42)聚集、抑制Aβ_(1-42)介导的神经毒性,与未修饰的柚皮苷相比,生物活性明显提高;该化合物能减少经Cu~(2+)-Aβ_(1-42)处理的PC12细胞中的ROS以及提高Cu~(2+)-Aβ_(1-42)处理的PC12细胞中SOD活性。这一研究工作显示,柚皮苷双席夫碱可能是一种具有潜在治疗阿尔茨海默病的药物。     

利用分子雷达研究吗啡对PC12细胞的作用机制

分子雷达是一种能够在分子层次对活细胞内生物过程进行实时、原位观察的探测技术。本文利用分子雷达研究了吗啡对PC12细胞的作用机制。实验结果表明，吗啡对PC12的作用具有时间和浓度依赖性，吗啡能引起细胞内[Ca^2 ]瞬时升高和波动，并且能导致细胞核浓缩和线粒体膜电势丧失。     

3-(2-萘基)-1-苯基-吡唑-4-甲醛腙衍生物的简便合成与表征

为寻找具有生物活性的先导化合物,以3-(2-萘基)-1-苯基-吡唑-4-甲醛为起始原料,在超声波辐射下经串联反应和"一锅法合成"两种途径制备了一系列新型的1-(3-β-萘基-1-苯基吡唑-4-亚甲基)-2-(4-芳基噻唑-2-基)-腙及其衍生物.与加热实验相比,在超声波辐射下可得到较好的产率.利用1H NMR,MS,IR谱和元素分析对产物进行了结构表征.初步的生物活性测试结果表明,化合物4b和4d对H2O2诱导的PC12细胞损伤具有明显的保护作用.     

栀子酰胺A-他克林杂合体的设计合成及其对6-羟多巴胺诱导的PC12细胞损伤的保护作用

设计合成了7个栀子酰胺A(GA)-他克林杂合物.首先以京尼平为原料,经5步反应合成10-栀子酰胺A醛(DG),总收率20.4%;再以邻氨基苯甲酸甲酯为原料,经4步反应合成N-末端氨基烷基他克林(TN-n),总收率50.0%~65.4%;随后,DG与TN-n在合适的还原剂作用下还原胺化获得目标化合物,收率37%~54%.所有目标产物的结构均由1H NMR,~(13)C NMR和ESI-MS确证.同时建立了六羟多巴胺(6-OHDA)诱导损伤的PC12细胞模型对系列杂合物进行生物活性评价.发现杂合物TNG-1和TNG-5具有比他克林、GA及其联合用药更好的神经保护作用,推测这两个杂合物可能具有比单体药物更好的类药性.     

Exploration of quinoxaline derivatives as antimicrobial and anticancer agents

Novel 2 and 3‐substituted quinoxaline derivatives were synthesized through various synthetic pathways, among which cyanoacetamide and cyanoacetohydrazide quinoxaline derivatives 4a‐c and 11a‐c , respectively, were synthesized. Furthermore, methoxy quinoxaline derivatives 3c and quinoxaline derivatives bearing substituted pyridines 6a,b , 12a,b , and 13a,b were designed to be synthesized. However, we have synthesized acrylohydrazide 5a,b and 7 /acrylamide derivatives, Schiff base analogues 14a‐f , pyrazole derivatives 15a‐e, amide derivatives 16a‐f , guanidine derivatives 16 g,h as well as, quinoxalin‐2‐methylallyl propionate derivative 14g . All the synthesized compounds were confirmed via spectral data and elemental analyses. Moreover, the newly synthesized compounds were evaluated for their antimicrobial activity (Gm +ve, Gm ?ve in comparison to Gentamycin a standard) and fungi (in comparison to Ketoconazole as a standard). Thus, compound 16b showed promising antimicrobial activity against B. subtilis, P. vulgaris, and S. mutants with values ranging from 20 to 27‐mm zone of inhibition. While compounds 5a , 14e,f, and 16a,c,d,g,h showed potent antimicrobial activity. Moreover, the National Cancer Institute (NCI) selected 20 compounds that were submitted for anticancer screening against 60 types of cancer cell lines. The most active compounds are 5b and 12a where compound 5b containing 2,4‐dichlorophenyl moiety at cyanoacetamide linkage of hydrazine quinoxaline backbone exerted significant growth inhibition activity against Leukemia MOLT‐4, Renal cancer UO‐31, and Breast cancer MCF‐7. In addition, compound 12a having 4,6‐diaminopyridinone side chain at position‐3 of quinoxaline nucleus exhibited remarkable anticancer activity against renal cancer UO‐31.     

Synthesis of Aminoalkyl-substituted Polymethoxychalcone Derivatives and Their Antiproliferative Activities Against Three Human Cancer Cell Lines

Two novel series of sixteen aminoalkyl-substituted polymethoxychalcone derivatives 2a-2h and 3a-3h were synthesized from 2'-hydroxy-3,4,5,4',6'-pentamethoxy chalcone(1) through extending alkoxy side chain at the 2'-position, and introducing amine hydrogen bond receptor at the end of the side chain. The structures of all the synthesized compounds were confirmed by ¹H NMR, ¹³C NMR and MS techniques. Furthermore, all the compounds were tested for antiproliferative activities in vitro against a panel of three human cell lines(HeLa, HCC1954 and SK-OV-3) via CCK-8 assay. The results show that all the target compounds exhibit antiproliferative activities against the three human cancer cells with IC₅₀ values of 4.62-48.21 μmol/L, except compound 2h against SK-OV-3 cells. Most of these compounds were more active when compared to the positive control cis-Platin.     

Monoterpene Indole Alkaloids from the Aerial Parts of Rhazya stricta Induce Cytotoxicity and Apoptosis in Human Adenocarcinoma Cells

Chromatographic investigation of the aerial parts of the Rhazya stricta (Apocynaceae) resulted in the isolation of two new monoterpene indole alkaloids, 6-nor-antirhine-N₁-methyl (1) and razyamide (2), along with six known compounds, eburenine (3), epi-rhazyaminine (4), rhazizine (5), 20-epi-sitsirikine (6), antirhine (7), and 16-epi-stemmadenine-N-oxide (8). The chemical structures were established by various spectroscopic experiments. Compounds 1–8 exhibited cytotoxic effects against three cancer cells with IC₅₀ values ranging between 5.1 ± 0.10 and 93.2 ± 9.73 µM against MCF-7; 5.1 ± 0.28 and 290.2 ± 7.50 µM against HepG2, and 3.1 ± 0.17 and 55.7 ± 4.29 µM against HeLa cells. Compound 2 showed the most potent cytotoxic effect against all cancer cell lines (MCF-7, HepG2 and HeLa with IC₅₀ values = 5.1 ± 0.10, 5.1 ± 0.28, and 3.1 ± 0.17 µM, respectively). Furthermore, compound 2 revealed a significant increase in the apoptotic cell population of MCF-7, HepG2, and HeLa cells, with 31.4 ± 0.2%, 29.2 ± 0.5%, and 34.9 ± 0.6%, respectively. Compound 2 decreased the percentage of the phagocytic pathway on HepG2 cells by 15.0 ± 0.1%. These findings can explain the antiproliferative effect of compound 2.     

Bioactive compounds from Selaginella involven Spring that protect PC-12 cells

Two new compounds named as 3b 12 16-trihydroxy-6 8 11 13-abietatrien（1） （8R 80S）-4 40 8-trihydroxyl-3 30-dimethoxyl-90-lignanolide（2） and a new natural product 4 40-dihydroxyl-3 30 5 50-dimethoxyldiphenyl diketone（3） were isolated from the whole herbs of Selaginella involven Spring.The structures were elucidated by spectroscopic analyses including UV,IR,1D,2D NMR and MS methods.Additionally,these three compounds exhibited potent protective effect against the injury of PC-12 cells induced by hypoxia/reoxygenation.     

Synthesis and cytotoxic activity of some new heterocycles incorporating cyclohepta[b]thiophene-3-carboxamide derivatives

A series of 2-amino-5,6,7,8-tetrahydro-4H-cyclohepta[b]thiophene-3-carboxamide-heterocyclic hybrids were synthesized, characterized and their cytotoxic potencies were assessed on four human cell lines. Cyanoacetamide derivative ( 5 ) was used as the key synthetic intermediate for the synthesis many derivatives in this study, derivatives 9 , 11 , 12 were formed by coupled compound 5 with different aryl/heteryl diazonium chlorides, Gewald reaction and Knoevenagel condensation were used for synthesis derivatives 13 , 14 , 16 by treated cyanoacetamide ( 5 ) with different reagents. In another route, compound 5 treated with phenyl isothiocyanate give thiocarbamoyl derivative ( 7 ) which used as intermediate underwent oxidative cyclization with different moieties to offer the corresponding thiazoles and thiophene 18 , 19 , 20 , 21 , respectively. in vitro cytotoxic activity of prepared compounds were tested against four human tumor cell lines. The result revealed that compound 11a displayed promising cytotoxic activity against HepG2, HCT-116, MCF-7, and PC3 cancer cell lines comparing to the positive control (Doxorubicin).     

Total synthesis of oxa-9-anthracyclines

Racemic 7-hydroxy-9-oxa-anthracyclinone (5a) has been synthetised in seven steps from quinizarin (6) and its resolution achieved after glycosylation with 3,4-di-O-acetyl-2-deoxy-L-fucose. Chiral pool syntheses of (8S)-8-hydroxymethyl-9-oxa-anthracyclinone (5b) and of (8S,10R) and (8S,10S)-8-hydroxymethyl-10-methyl-9-oxa-anthracyclinones (5c and 5d) have been achieved using (R)-2,3-O-isopropylideneglyceraldehyde (12) and leucoquinizarin (13) as starting materials. Glycosylation of aglycones 5b-5d by either 3,4-di-O-acetyl-2-deoxy-L-fucose or various 3-amino-2,3,6-trideoxy-L-hexoses yielded the corresponding anthracyclines. The synthetic glycosides do not show significant cytotoxic activity at a concentration of 1 microgram/ml against L 1210 cells.     

MiodesinTM Positively Modulates the Immune Response in Endometrial and Vaginal Cells

Endometriosis presents high prevalence and its physiopathology involves hyperactivation of endometrial and vaginal cells, especially by bacteria. The disease has no cure and therapies aiming to inhibit its development are highly desirable. Therefore, this study investigated whether Miodesin^TM (10 µg/mL = IC₈₀; 200 µg/mL = IC₅₀), a natural compound constituted by Uncaria tomentosa, Endopleura uchi, and astaxanthin, could exert anti-inflammatory and anti-proliferative effects against Lipopolysaccharides (LPS) stimulation in endometrial and Candida albicans vaginal cell lines. VK2 E6/E7 (vaginal) and KLE (epithelial) cell lines were stimulated with Candida albicans (1 × 10⁷ to 5 × 10⁷/mL) and LPS (1 μg/mL), respectively. Miodesin^TM inhibited mRNA expression for Nuclear factor kappa B (NF-κB), ciclo-oxigenase 1 (COX-1), and phospholipase A2 (PLA2), beyond the C–C motif chemokine ligand 2 (CCL2), CCL3, and CCL5 in VK2 E6/E7 cells (p < 0.05). In addition, the inhibitory effects of both doses of Miodesin^TM (10 µg/mL and 200 µg/mL) resulted in reduced secretion of interleukin-1β (IL-1β), IL-6, IL-8, tumor necrosis factor α (TNF-α) (24 h, 48 h, and 72 h) and CCL2, CCL3, and CLL5 (p < 0.05) by VK2 E6/E7 cells. In the same way, COX-1 Miodesin^TM inhibited LPS-induced hyperactivation of KLE cells, as demonstrated by reduced secretion of IL-1β, IL-6, IL-8, TNF-α (24 h, 48 h, and 72 h) and CCL2, CCL3, and CLL5 (p < 0.05). Furthermore, Miodesin^TM also inhibited mRNA expression and secretion of matrix metalloproteinase-2 (MMP-2), MMP-9, and vascular endothelial growth factor (VEGF), which are key regulators of invasion of endometrial cells. Thus, the study concludes that Miodesin^TM presents beneficial effects in the context of endometriosis, positively affecting the inflammatory and proliferative response.     

Design,Cytotoxicity and Antiproliferative Activity of 4-Amino-5-methyl-thieno[2,3-d]pyrimidine-6-carboxylates against MFC-7 and MDA-MB-231 Breast Cancer Cell Lines

Novel 4-amino-thieno[2,3-d]pyrimidine-6-carboxylates substituted at the second position were prepared by cyclocondensation of 2-amino-3-cyano-thiophene and aryl nitriles in an acidic medium. The design of the target compounds was based on structural optimization. The derivatives thus obtained were tested in vitro against human and mouse cell lines. The examination of the compound effects on BLAB 3T3 and MFC-10A cells showed that they are safe, making them suitable for subsequent experiments to establish their antitumor activity. The photoirritancy factor of the compounds was calculated. Using the MTT test, the antiproliferative activity to MCF-10A, MCF-7 and MDA-MB-231 cell lines was estimated. The best antiproliferative effect in respect to the MCF-7 cell line revealed compound 2 with IC₅₀ 4.3 ± 0.11 µg/mL (0.013 µM). The highest selective index with respect to MCF-7 cells was shown by compound 3 (SI = 19.3), and to MDA-MB-231 cells by compound 2 (SI = 3.7). Based on energy analysis, the most stable conformers were selected and optimized by means of density functional theory (DFT). Ligand efficiency, ligand lipophilicity efficiency and the physicochemical parameters of the target 4-amino-thienopyrimidines were determined. The data obtained indicated that the lead compound among the tested substances is compound 2.     

Target Design of Novel Histone Deacetylase 6 Selective Inhibitors with 2-Mercaptoquinazolinone as the Cap Moiety

Epigenetic alterations found in all human cancers are promising targets for anticancer therapy. In this sense, histone deacetylase inhibitors (HDACIs) are interesting anticancer agents that play an important role in the epigenetic regulation of cancer cells. Here, we report 15 novel hydroxamic acid-based histone deacetylase inhibitors with quinazolinone core structures. Five compounds exhibited antiproliferative activity with IC₅₀ values of 3.4–37.8 µM. Compound 8 with a 2-mercaptoquinazolinone cap moiety displayed the highest antiproliferative efficacy against MCF-7 cells. For the HDAC6 target selectivity study, compound 8 displayed an IC₅₀ value of 2.3 µM, which is 29.3 times higher than those of HDAC3, HDAC4, HDAC8, and HDAC11. Western blot assay proved that compound 8 strongly inhibited tubulin acetylation, a substrate of HDAC6. Compound 8 also displayed stronger inhibition activity against HDAC11 than the control drug Belinostat. The inhibitory mechanism of action of compound 8 on HDAC enzymes was then explored using molecular docking study. The data revealed a high binding affinity (−7.92 kcal/mol) of compound 8 toward HDAC6. In addition, dock pose analysis also proved that compound 8 might serve as a potent inhibitor of HDAC11.     

Synthesis of Novel Methyl 3-(hetero)arylthieno[3,2-b]pyridine-2-carboxylates and Antitumor Activity Evaluation: Studies In Vitro and In Ovo Grafts of Chick Chorioallantoic Membrane (CAM) with a Triple Negative Breast Cancer Cell Line

A series of novel functionalized methyl 3-(hetero)arylthieno[3,2-b]pyridine-2-carboxylates 2a–2h were synthesized by C-C Pd-catalyzed Suzuki-Miyaura cross-coupling of methyl 3-bromothieno[3,2-b]pyridine-2-carboxylate with (hetero)aryl pinacol boranes, trifluoro potassium boronate salts or boronic acids. Their antitumoral potential was evaluated in two triple negative breast cancer (TNBC) cell lines—MDA-MB-231 and MDA-MB-468, by sulforhodamine B assay. Their effects on the non-tumorigenic MCF-12A cells were also evaluated. The results demonstrated that three compounds caused growth inhibition in both TNBC cell lines, with little or no effect against the non-tumorigenic cells. The most promising compound was further studied concerning possible effects on cell viability (by trypan blue exclusion assay), cell proliferation (by bromodeoxyuridine assay) and cell cycle profile (by flow cytometry). The results demonstrated that the GI₅₀ concentration of compound 2e (13 μM) caused a decreased in MDA-MB-231 cell number, which was correlated with a decreased in the % of proliferating cells. Moreover, this compound increased G0/G1 phase and decreased S phases, when compared to control cells (although was not statistic significant). Interestingly, compound 2e also reduced tumor size using an in ovo CAM (chick chorioallantoic membrane) model. This work highlights the potential antitumor effect of a novel methyl 3-arylthieno[3,2-b]pyridine-2-carboxylate derivative.