IOP-lowering effect of isoquinoline-5-sulfonamide compounds in ocular normotensive monkeys

Rho-associated coiled coil-formed protein kinase (ROCK) inhibitors are under development as a new class of antiglaucoma agents. Based on the potent ROCK inhibitor H-1152, previously developed by us, we explored the possibility of related compounds as antiglaucoma agents and synthesized seven types of H-1152-inspired isoquinoline-5-sulfonamide compounds (H-0103–H-0107, H-1001, H-1005). Although all of these compounds potently inhibited ROCK (IC₅₀ = 18–48 nM), only H-0104 and H-0106 exerted strong intraocular pressure (IOP)-lowering effects into the eyes of monkeys. These results suggested the possibility that there is no direct relationship between ROCK inhibition and IOP-lowering effects, indicating that the initial screening of compounds based on ROCK inhibitory activity may be an unsuitable strategy for developing antiglaucoma agents with potent IOP-lowering effects.     

Environmental restoration effects of Ranunculus sceleratus L. in a eutrophic sewage system

This study investigated the environmental restoration effects of Ranunculus sceleratus in a sewage system microcosm trial, including the removal of pollutants and algal inhibition. We compared the removal of pollutants by R. sceleratus in a eutrophic sewage system in the presence and the absence of algae. The rate of removal without algae was 16.2–20.5% of that with algae. ${\text{NH}}_4^{+}–\text{N}$ was removed most readily by R. sceleratus. The effects of R. sceleratus on the growth of Microcystis aeruginosa were also investigated in two allelopathic modes. The level of algal inhibition after the addition of an extract of Ranunculus scleratus was 57.1–78.9% greater than that in a co-culture test. To understand the role of allelopathy interference with algal development, we also determined the total flavonoid contents of plants, which ranged from 3.57 g to 20.19 g per plant. The cell density of Microcystis aeruginosa was negatively correlated with the total flavonoids in R. sceleratus, although aquatic macrophytes may contain other allelochemicals involved with algal inhibition in addition to flavonoid compounds. The environmental effects of R. sceleratus were significantly correlated with its growth stage (or water retention time), plant height, and biomass. This study suggests that R. sceleratus has potential for the low-effort and sustainable management of freshwaters, particularly the removal of nutrient pollutants and the reduction of excessive algal growth, which may be attributable to allelochemicals such as flavonoids. The in situ environmental restoration effects of R. sceleratus require further investigation at the ecosystem level.     

2,2′-Dihydroxybenzophenones and their carbonyl N-analogues as inhibitor scaffolds for MDR-involved human glutathione transferase isoenzyme A1-1

The MDR-involved human GSTA1-1, an important isoenzyme overexpressed in several tumors leading to chemotherapeutic-resistant tumour cells, has been targeted by 2,2′-dihydroxybenzophenones and some of their carbonyl N-analogues, as its potential inhibitors. A structure-based library of the latter was built-up by a nucleophilic cleavage of suitably substituted xanthones to 2,2′-dihydroxy-benzophenones (5–9) and subsequent formation of their N-derivatives (oximes 11–13 and N-acyl hydrazones 14–16). Screening against hGSTA1-1 led to benzophenones 6 and 8, and hydrazones 14 and 16, having the highest inhibition potency (IC₅₀ values in the range 0.18 ± 0.02 to 1.77 ± 0.10 μM). Enzyme inhibition kinetics, molecular modeling and docking studies showed that they interact primarily at the CDNB-binding catalytic site of the enzyme. In addition, the results from cytotoxicity studies with human colon adenocarcinoma cells showed low LC₅₀ values for benzophenone 6 and its N-acyl hydrazone analogue 14 (31.4 ± 0.4 μM and 87 ± 1.9 μM, respectively), in addition to the strong enzyme inhibition profile (IC₅₀₍₆₎ = 1,77 ± 0.10 μM; IC₅₀₍₁₄₎ = 0.33 ± 0.05 μM). These structures may serve as leads for the design of new potent mono- and bi-functional inhibitors and pro-drugs against human GTSs.     

Oxidation of cyanobenzocycloheptatrienes: Synthesis,photooxygenation reaction and carbonic anhydrase isoenzymes inhibition properties of some new benzotropone derivatives

The oxidation of some cyanocycloheptatrienes with CrO₃ and pyridine was investigated and a few new nitrile functionalised benzotropone derivatives were obtained. Photooxygenation reaction of these products was also studied. The structures of the formed products were determined on the basis of NMR spectroscopy and the formation mechanism of unusual products was discussed. Human carbonic anhydrase isoenzymes I, and II (hCA I and hCA II) inhibition properties of nitrile functionalized new benzotropone derivatives were also studied. Both CA isozymes were inhibited in the low micromolar range by these nitrile functionalized benzotropone analogues. The newly synthesized benzotropone derivatives showed inhibition constants in the sub-micromolar range (2.51–4.06 μM). The best hCA I inhibition was observed in 5H-benzocycloheptene-7-carbonitrile (K_i: 2.88 ± 0.86 μM). On the other hand, 5-oxo-5H-benzocycloheptatriene-7-carbonitrile showed the powerful inhibitory effect against hCA II (K_i: 2.51 ± 0.34 μM).     

Synthesis, β-haematin inhibition,and in vitro antimalarial testing of isocryptolepine analogues: SAR study of indolo[3,2-c]quinolines with various substituents at C2, C6, and N11

A series of indolo[3,2-c]quinolines were synthesized by modifying the side chains of the ω-aminoalkylamines at the C6 position and introducing substituents at the C2 position, such as F, Cl, Br, Me, MeO and NO₂, and a methyl group at the N11 position for an SAR study. The in vitro antiplasmodial activities of the derivative agents against two different strains (CQS: NF54 and CQR: K1) and the cytotoxic activity against normal L6 cells were evaluated. The test results showed that compounds 6k and 6l containing the branched methyl groups of 3-aminopropylamino at C6 with a Cl atom at C2 exhibited a very low cytotoxicity with IC₅₀ values above 4000 nM, high antimalarial activities with IC₅₀ values of about 11 nM for CQS (NF54), IC₅₀ values of about 17 nM for CQR (K1), and RI resistance indices of 1.6. Furthermore, the compounds were tested for β-haematic inhibition, and QSAR revealed an interesting linear correlation between the biological activity of CQS (NF54) and three contributing factors, namely solubility, hydrophilic surface area, and β-haematin inhibition for this series. In vivo testing of 6l showed a reduction in parasitaemia on day 4 with an activity of 38%.     

Viral Cyclin promotes KSHV-induced cellular transformation and tumorigenesis by overriding contact inhibition

Kaposi sarcoma-associated herpesvirus (KSHV) is a tumor virus encoding several proto-oncogenes. However, the roles of these viral genes in KSHV-induced tumorigenesis have not been defined. In this study, we used a recently developed model of KSHV-induced cellular transformation and tumorigenesis combining with a reverse genetic system to examine the role of a KSHV latent gene vCyclin (ORF72), a cellular Cyclin D2 homolog, in KSHV-induced oncogenesis. Deletion of vCyclin did not affect cell proliferation and cell cycle progression at a low-density condition, when cells were at an active proliferation state. However, vCyclin mutant cells were contact-inhibited and arrested at G₁ phase at a high-density condition. As a result, vCyclin mutant cells formed less and smaller colonies in soft agar assay. Nude mice inoculated with vCyclin mutant cells had reduced tumor incidence and extended tumor latency and survival compared with mice inoculated with wild-type (WT) virus-infected cells. WT but not mutant virus effectively induced Cyclin-dependent kinase inhibitor p27/Kip1 Ser10 phosphorylation and cytoplasmic relocalization. shRNA knockdown of p27 released the blockage of the mutant cells from cell cycle arrest at G₁ phase at a high-density condition. Together, these results indicate that vCyclin primarily functions to enhance cellular transformation and tumorigenesis by promoting cell cycle progression and cell proliferation at a contact-inhibited condition.     

高压电场不可逆电穿孔诱发A549肺癌细胞凋亡的实验研究

目的：不可逆电穿孔是治疗肿瘤的新兴技术,本文探讨高压电场引起的不可逆电穿孔诱发A549肺癌细胞凋亡的特点。方法：选择处于生长周期的A549细胞,共分为A—G7个组进行研究,其中A组为不施加电场的空白对照组,B-G组为实验组,B组施加500V／cm强度高压电场,G组施加1750V／cm的高压电场,BG组之间各组的高压电场强度间隔为250V／cm。采取细胞抑制实验、不可逆电穿孔示踪实验、细胞凋亡实验,检验A549细胞细胞凋亡与电场强度的关系。结果：①各实验组与对照组、各实验组之间的细胞抑制率,均存在显著性差异（P〈0．05）;②电场强度≥1000V／cm时,细胞不可逆电穿孔率明显增加,有统计学意义（P〈0．05）;电场强度≥1500V／cm时,细胞不可逆电穿孔率增加不明显,无统计学意义（P〉0．05）;③电场强度≥1250V／cm时,细胞早期凋亡率明显增加,有统计学意义（P〈0．05）。结论：高压电场不可逆电穿孔诱发A549肺癌细胞发生早期凋亡的强度为1250V／cm,发生晚期凋亡的强度为1500V／cm,且凋亡率随着电场强度的增加持续升高。这对于高压电场不可逆电穿孔效应引起的肿瘤细胞凋亡机制的研究具有重要意义。     

骆驼蓬脂转移蛋白基因真核表达载体的构建及其体内外抗瘤效应的研究

目的:构建骆驼蓬脂转移蛋白(lipid transfer protein from Peganum harmala,PhLTP)基因真核表达质粒,并探讨其对黑色素瘤B16细胞在体内外的抗肿瘤作用。方法:将PhLTP基因亚克隆至pcDNA3.1上,获得重组质粒pcDNA3.1-PhLTP;用脂质体转染法将重组质粒及空载体外转染B16细胞,MTT检测其对B16细胞生长的影响。建立B16荷瘤小鼠模型,设重组质粒(pcDNA3.1-PhLTP)、空载(pcDNA3.1)、生理盐水和阳性药物(CTX)组,分别处理小鼠后测量各组肿瘤体积并称瘤重,计算抑瘤率。光镜观察鼠脾、肝等组织变化;免疫组织化学法检测各瘤体中PhLTP、血管内皮生长因子(VEGF)及碱性成纤维细胞生长因子(bFGF)的表达。结果:pcDNA3.1-PhLTP转染B16细胞72 h后,细胞增殖能力明显受到抑制(P0.01)。注射pcDNA3.1-PhLTP组的小鼠肿瘤生长速度明显减慢,肿瘤体积小于空载和生理盐水组(P0.05)。显微镜下可见重组质粒组肿瘤细胞有不同程度的点、片状坏死,而肝、肺等无明显病理损伤。重组质粒组肿瘤组织中有PhLTP蛋白的表达,且VEGF和bFGF的阳性表达指数都低于空载和生理盐水组(P0.01)。结论:成功构建了重组表达质粒pcDNA3.1-PhLTP,体内外实验结果显示其能有效地抑制B16细胞的生长,预示了该重组质粒在治疗黑色素瘤中的潜在应用价值。     

Haloperidol treatment at pre-exposure phase reduces the disturbance of latent inhibition in rats with neonatal ventral hippocampus lesions

Animals with neonatal ventral hippocampal lesions develop during or after adolescence abnormal behaviors related to schizophrenia such as anxiety and latent inhibition disruption. The aim of this study was to test whether haloperidol injection prior to pre-exposure session in the latent inhibition test would facilitate latent inhibition.