Evodiamine-inspired dual inhibitors of histone deacetylase 1 (HDAC1) and topoisomerase 2 (TOP2) with potent antitumor activity

A great challenge in multi-targeting drug discovery is to identify drug-like lead compounds with therapeutic advantages over single target inhibitors and drug combinations. Inspired by our previous efforts in designing antitumor evodiamine derivatives, herein selective histone deacetylase 1 (HDAC1) and topoisomerase 2 (TOP2) dual inhibitors were successfully identified, which showed potent in vitro and in vivo antitumor potency. Particularly, compound 30a was orally active and possessed excellent in vivo antitumor activity in the HCT116 xenograft model (TGI = 75.2%, 150 mg/kg, p.o.) without significant toxicity, which was more potent than HDAC inhibitor vorinostat, TOP inhibitor evodiamine and their combination. Taken together, this study highlights the therapeutic advantages of evodiamine-based HDAC1/TOP2 dual inhibitors and provides valuable leads for the development of novel multi-targeting antitumor agents.     

离子选择电极电位法研究S2O8^2—与I^—反应动力学

用离子选择电极电位法对S_2O_8~(2-)-Ⅰ~-体系反应动力学问题进行了探讨,研究了离子强度对反应速度的影响。结果表明:离子选择电极电位法可用于此体系的动力学研究,并且得到满意的结果;提出了该电极适用的浓度条件;得出生成CuI~+步骤为催化反应决定步骤的新结论;随着离子强度的增加dE/dt值增大,即反应速度加快。     

离子碘和分子碘过量对甲状腺功能与形态影响

目的　比较离子碘和分子碘过量对非碘缺乏Wistar大鼠甲状腺功能和形态的影响及差别。方法　固相免疫放射分析法测定实验大鼠血清TSH ,放射免疫分析法测定血清TT4 、TT3 。砷铈分光光度法测定尿碘。光、电镜下观察甲状腺内形态学变化 ,用图像分析仪测量滤泡上皮细胞高度和滤泡腔面积。结果　补充 840 μg碘 /L 90d时 ,可使血清TSH值增高 ,但是未见显著性差异。血清TT4 明显高于双蒸馏水 (DDW )组 ,P <0 0 5,但同剂量离子碘和分子碘比较 ,血清TT4 值无显著性差异。补碘组光、电镜下可见滤泡腔面积增大 ,滤泡上皮细胞变扁 ,细胞核染色深 ,滤泡融合破裂 ,巨滤泡形成 ,毛细血管减少。超微结构显示甲状腺滤泡上皮细胞内质网扩张 ,次级溶酶体增多 ,微绒毛减少 ,染色体浓集。与DDW组相比 ,滤泡上皮细胞高度明显降低 ,滤泡腔面积增大 ,P均 <0 0 5。相同剂量离子碘和分子碘比较无显著性差异。结论　碘摄入浓度 >840 μg/L时 ,会对大多部分甲状腺滤泡上皮细胞产生抑制和破坏 ,使血清TT4 值明显增高 ,离子碘和分子碘对甲状腺影响无明显区别     

Na+/H+ exchange regulates intracellular pH of rat gastric surface cells in vivo

Intracellular pH (pH_i) and viability of gastric surface cells of the rat stomach in response to luminal acidification, and the role of Na⁺/H⁺ exchange in maintaining pH_i homeostasis were studied in vivo using a fluorescent microscopic technique. pH_i was measured during superfusion with buffers of pH 1.2–7.4. When the pH of the superfusate was 7.4, baseline pH_i was unchanged. Superfusion with pH 3 buffer rapidly decreased pH_i to 6.7, with subsequent recovery to baseline pH_i within 15 min despite continuing acid exposure. Superfusion with buffers of pH 1.7 and 1.2 decreased pH_i continuously to below 6.2 with no recovery observed. Despite the relentless decline in pH_i during superfusion with pH-1.2 and –1.7 solutions, over 75% of the surface cells were still viable, as measured by exclusion of the vital dye propidium iodide. We then examined the role of Na⁺/H⁺ exchange in the regulation of pH_i. Superfusion with amiloride did not affect recovery of pH_i from intracellular acidification induced by a NH₄Cl prepulse. Exposure to the potent, lipophilic Na⁺/H⁺ exchange inhibitor 5-(N,N-hexaniethylene)-amiloride (HMA), either in the superfusate or by close arterial perfusion, decreased baseline pH_i from 7.1 to 6.8. Close arterial perfusion of HMA additionally attenuated the recovery of pH_i to baseline during superfusion with pH 3 buffer. We conclude that luminal protons permeate into the cytoplasm of gastric surface cells, where they are eliminated by an Na⁺/H⁺ exchanger, most probably localized to the basolateral membrane.     

Characterization of a monoclonal anti-Bw4 antibody (Tü109): Evidence for similar epitopes on the Bw4 and Bw6 antigens

The production and serologic, as well as immunochemical properties of a cytotoxic murine IgG monoclonal antibody (Tü109) that precipitates HLA-class I molecules, are described. In the microcytotoxicity assay Tü109 supernatant was demonstrated on a panel of 424 HLA-ABC, -DR, -DQ, -MT typed normal Caucasian blood donors to define an epitope on HLA-B locus molecules in great association with the supertypic specificity Bw4. Reactivity of supernatant showed MHC linked inheritance of the Tü109 determinant and discriminated the HLA-Bw4/Bw6 associated HLA-B locus split antigens. Weak or lack of binding on lymphocytes from some HLA-Bw4 heterozygous individuals, particularly typing for HLA-Bw44, appeared to be due to qualitative and/or quantitative variations of HLA-B locus molecules on the cell surface. With Tü109 ascites fluid, however, extra-reactivity on all HLA-Bw6⁺ cells was demonstrated. Preferential binding of supernatant to HLA-Bw4, but reactivity of ascites fluid with HLA-Bw6⁺ molecules in addition, was furthermore confirmed by IEF analysis of antigens immunoprecipitated with Tü109 from cell lysates. Thus the antibody may help to analyze the evolutionary relationship of the diallelic specificities Bw4 and Bw6.     

Effect of potassium iodide and perchlorate on the secretion of thyroid hormones

The effect of potassium iodide and perchlorate was studied on parameters characterizing the secretion of thyroid hormones: the cyclic AMP level in the thyroid gland tissue and the number of intracellular colloid drops during stimulation with thyroid hormones. Excess of iodide was shown to depress these indices whereas perchlorate prevented the inhibitory effect of iodide. The results support the view that the adenylate-cyclase system of the thyroid gland is sensitive to the organic iodine concentration. Excess of iodide evidently reduces the ability of perchlorate to affect its concentration in the gland, so that the process of organification of iodine and the secretion of thyroid hormones are maintained at the optimal level.Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 12, pp. 653–655, December, 1978.     

Chimaeric anti-CD4 monoclonal antibody cross-linked by monocyte Fcγ receptor mediates apoptosis of human CD4 lymphocytes

Previous studies have shown that murine anti-CD4 monoclonal antibody, cross-linked by rabbit anti-mouse immunoglobulin, could mediate apoptosis of murine CD4⁺ lymphocytes when they were stimulated by T cell receptor antibody. In this study, we have shown that the murine anti-CD4 monoclonal antibody, OKT4, can induce apoptosis in human CD4⁺ T cells stimulated by the recall antigen tuberculin purified protein derivative (PPD) only when cross-linked by rabbit anti-mouse immunoglobulin. The chimeric anti-CD4 monoclonal antibody, cM-T412 whose Fc fragment is human, was able to cause apoptosis without cross-linking by a second antibody. Similarly, abolition of PPD-induced proliferation of peripheral blood mononuclear cells by cM-T412 did not require cross-linking with rabbit anti-human immunoglobulin. Inhibition of proliferation by cM-T412 could be reduced by pre-treating monocytes with heat-aggregated human IgG. This suggested that monocyte Fcγ receptors might be cross-linking the human Fc of cM-T412. Propidium iodide staining together with immunofluorescence showed that the apoptotic cells were indeed CD4⁺ lymphocytes. It is proposed that during treatment with cM-T412 in autoimmune disease such as rheumatoid arthritis, cM-T412-coated CD4 T cells, when they are subsequently stimulated by the unknown arthritogenic antigen, may undergo apoptotic cell death through cross-linking of cM-T412 on Fey receptor-positive cells within the joint.     

Isolation and partial characterization of the tegumental outer membrane of schistosomula of Schistosoma mansoni

Separation of the external membranes from freshly converted mechanical schistosomula of Schistosoma mansoni was achieved by osmotic shock under hypertonic conditions, followed by mechanical shearing and ultracentrifugation. Prior to treatment, the schistosomula were surface labeled by introduction of N-DNP-epsilon-aminocaproylphosphatidylethanolamine molecules into their lipid bilayer followed by anti-DNP antibodies and stained with either 125I-protein-A or ferritin labeled secondary anti-DNP antibodies. This label provided a membrane marker by which the purity of the preparation could be assessed at each stage. Fluorescence staining with FITC-conjugated secondary antibodies prior to treatment revealed that the homogeneously stained membrane of the intact schistosomula became swollen and ruptured after the osmotic shock. The isolated membrane pellet was intensely fluorescent. Electron microscopical examination revealed mostly vesicles, some of them with organized multilayer assembly. The vesicles were ferritin labeled, indicating that they originated from the outer surface membrane of the schistosomula. A 100 fold enrichment in the alkaline phosphatase activity and about 300 fold enrichment in acetylcholinesterase activity in the membrane preparations, as compared to the intact schistosomula, was found. The isolated tegument was analyzed by SDS-polyacrylamide gel electrophoresis. The pattern obtained showed three major bands, of molecular weights 69 000, 45 000 and 12 000 alongside with a large number of minor bands. Immunoprecipitation of the isolated 125I-labeled membrane antigens with antisera from chronically infected mice revealed these three major bands together with three other bands of molecular weight 38 000, 23 000 and 16 000.     

碘化钠对人甲状腺上皮细胞TNF-α、IL-1β分泌的影响

为研究碘化钠 (NaI)对人甲状腺上皮细胞 (TEC )分泌细胞因子TNF α和IL 1β的影响 ,以探讨碘在GD病 (GD )发病中的可能机制 ,取手术切除的GD病患者甲状腺组织和甲状腺腺瘤患者瘤旁正常甲状腺组织 ,进行细胞培养。以不同浓度的NaI (10 8～ 10 3 mol/L )刺激单层培养的甲状腺细胞 ,采用放射免疫测定技术测定刺激前后甲状腺细胞培养上清液中细胞因子TNF α和IL 1β的含量。同时以透射电镜观察NaI刺激后甲状腺细胞的形态学改变。结果 :(1)正常甲状腺细胞能分泌少量的TNF α和IL 1β。GD的甲状腺细胞TNF α和IL 1β的分泌量与正常TEC相比显著增加 (P <0 0 1) ;(2 )当NaI浓度为 10 8～10 3 mol/L时 ,正常人甲状腺细胞TNF α、IL 1β的分泌量与 0mol/L组相比无显著性差异 (P >0 0 5 ) ;透射电镜示正常甲状腺细胞无损伤型改变 ;(3)当NaI浓度为 10 6～ 10 3 mol/L时 ,GD甲状腺细胞TNF α的分泌量显著增加 (P <0 0 5 ) ;当NaI浓度为 10 8～ 10 3 mol/L时 ,GD甲状腺细胞IL 1β的分泌量显著增加 (P <0 0 5 )。NaI浓度超过 10 6mol/L时 ,透射电镜示GD甲状腺细胞出现损伤型改变。表明高浓度的碘不仅造成GD甲状腺细胞的直接损伤 ,而且可以增加甲状腺细胞细胞因子TNF α和IL 1β的分泌 ,特别是在GD甲状腺细     

碘化钾、碘酸钾对缺碘大鼠甲状腺形态与作用的实验研究

目的：研究碘化钾、碘酸钾对大鼠甲状腺形态学结构的影响。方法：将Wistar大鼠随机分为4组,低碘组（LI）、正常组（NI）、碘化钾组（KI）、碘酸钾组（KIO3）,3个月后观察形态学变化,并做体视学分析,获得定量参数后进行统计学处理。结果：LI组滤泡及滤泡腔的平均体积VQ、平均表面积SQ、体积密度VV、表面积密度SV均明显小于NI组,而数密度NV及比表面积S/V均明显高于NI组;经治疗3个月后,KI组和KIO3组的上述指标均有明显恢复,但尚未完全恢复正常。KI组和KIO3组之间无显著性差异。结论：低碘可致大鼠甲状腺形态典型的小滤泡增生性改变,而KI和KIO3在治疗缺碘性甲状腺肿方面均有很好的疗效,且在本实验期间两者的疗效无显著性差异。