C-terminal trans,trans-muconic acid ethyl ester partial retro-inverso pseudopeptides as proteasome inhibitors

The development of specific inhibitors of the proteasome represents an important opportunity for new drug therapies. The central role of the multicatalytic complex in the intracellular proteolysis mediated by ubiquitin-proteasome pathway goes to discovery many molecules able to selectively inhibits enzymatic active subsites. Now, we report synthesis and activity of a new partial retro-inverso oligopseudopeptide derivatives bearing a trans,trans-muconic acid ethyl ester pharmacophoric unit at the C-terminal. Some analogues selectively inhibited in µM range the caspase-like (C-L) activity in the β1 subunit of the proteasome.     

植物类Caspase及其调控细胞程序性死亡的研究进展

植物细胞程序性死亡（PCD）在植物生长发育和逆境适应中发挥重要作用。半胱氨酸蛋白酶（caspase）调控动物PcD的启动、执行及信号转导。通过人工合成底物、动物caspase抑制剂等方法已证实在植物中存在类caspase,可分为metacas．pases、VPEs（vacuolar processing enzymes）和saspases等。本文综述了植物类caspase的种类、结构、定位、功能及其调控PCD的研究进展,提出植物PCD中类caspase作用的调控途径,为深入研究植物PCD提供参考。     

植物细胞程序性死亡中的类胱天蛋白酶研究进展

胱天蛋白酶(caspases)在动物细胞程序性死亡(programmed cell death,PCD)的起始、执行以及信号转导阶段起着关键作用,目前在植物中也发现有类胱天蛋白酶(caspase-like proteases,CLPs)的存在,并确认液泡加工酶(VPEs)、metacaspases和丝氨酸内肽酶(sapases)具有CLPs的作用,并证实CLPs参与植物的生长发育、抗病性及胁迫诱导的细胞程序性死亡等.本文对植物CLPs活性、生化结构及生理作用等方面的研究进展进行综述,并与动物caspases进行比较,为今后CLPs活性调节、作用方式及其在植物细胞程序性死亡中的作用等方面的研究提供参考.     

植物细胞程序性死亡研究进展

植物细胞死亡分为坏死和程序性死亡。细胞程序性死亡是具有信号或一系列分子参与,并且由细胞内在的死亡程序介导的有序过程。它在植物生长发育和抵御外界胁迫中具有重要作用。简要介绍了植物PCD的特征,对植物PCD中的信号分子和类caspase的作用等进行了综述,并对植物PCD存在的问题进行分析和展望,为深入研究植物PCD提供参考。     

Design,synthesis, in vitro and in vivo evaluation,and structure-activity relationship (SAR) discussion of novel dipeptidyl boronic acid proteasome inhibitors as orally available anti-cancer agents for the treatment of multiple myeloma and mechanism studies

A series of novel dipeptidyl boronic acid inhibitors of 20S proteasome were designed and synthesized. Aliphatic groups at R¹ position were designed for the first time to fully understand the SAR (structure–activity relationship). Among the screened compounds, novel inhibitor 5c inhibited the CT-L (chymotrypsin-like) activity with IC₅₀ of 8.21?nM and the MM (multiple myeloma) cells RPMI8226, U266B and ARH77 proliferations with the IC₅₀ of 8.99, 6.75 and 9.10?nM, respectively, which showed similar in vitro activities compared with the compound MLN2238 (biologically active form of marketed MLN9708). To investigate the oral availability, compound 5c was esterified to its prodrug 6a with the enzymatic IC₅₀ of 6.74?nM and RPMI8226, U266B and ARH77 cell proliferations IC₅₀ of 2.59, 4.32 and 3.68?nM, respectively. Furthermore, prodrug 6a exhibited good pharmacokinetic properties with oral bioavailability of 24.9%, similar with MLN9708 (27.8%). Moreover, compound 6a showed good microsomal stabilities and displayed stronger in vivo anticancer efficacy than MLN9708 in the human ARH77 xenograft mouse model. Finally, cell cycle results showed that compound 6a had a significant inhibitory effect on CT-L and inhibited cell cycle progression at the G2M stage.     

Caspase-like enzymatic activity and the ascorbate-glutathione cycle participate in salt stress tolerance of maize conferred by exogenously applied nitric oxide

Salinity stress causes ionic stress (mainly from high Na⁺ and Cl^- levels) and osmotic stress (as a result of inhibition of water uptake by roots and amplified water loss from plant tissue), resulting in cell death and inhibition of growth and ultimately adversely reducing crop productivity. In this report, changes in root nitric oxide content, shoot and root biomass, root H₂O₂ content, root lipid peroxidation, root cell death, root caspase-like enzymatic activity, root antioxidant enzymatic activity and root ascorbate and glutathione contents/redox states were investigated in maize (Zea mays L. cv Silverking) after long-term (21 d) salt stress (150 mM NaCl) with or without exogenously applied nitric oxide generated from the nitric oxide donor 2,2′-(Hydroxynitrosohydrazano)bis-ethane. In addition to reduced shoot and root biomass, salt stress increased the nitric oxide and H₂O₂ contents in the maize roots and resulted in elevated lipid peroxidation, caspase-like activity and cell death in the roots. Altered antioxidant enzymatic activities, along with changes in ascorbate and glutathione contents/redox status were observed in the roots in response to salt stress. The detrimental effects of salt stress in the roots were reversed by exogenously applied nitric oxide. These results demonstrate that exogenously applied nitric oxide confers salt stress tolerance in maize by reducing salt stress-induced oxidative stress and caspase-like activity through a process that limits accumulation of reactive oxygen species via enhanced antioxidant enzymatic activity.     

PGPR regulate caspase-like activity,programmed cell death,and antioxidant enzyme activity in paddy under salinity

The response of two root associated bacteria Pseudomonas pseudoalcaligenes and Bacillus pumilus were studied in the (salt-sensitive) rice GJ17 cultivar to salinity under controlled environmental growth conditions for protection of plant from adverse effect of salinity. Salinity affects the growth of salt-sensitive cultivar, but inoculation of plant growth promoting rhizobacteria (PGPR) reduces the harmful effect of salinity. The present study states that PGPR helps to reduce lipid peroxidation and superoxide dismutase activity in salt-sensitive GJ17 cultivar under salinity and play an important role in the growth regulation for positive adaptation of plants to salt stress. This study shows that inoculation of paddy (Oryza sativa) with such bacteria could provide salt-tolerant ability by reducing the toxicity of reactive oxygen species by reducing plant cell membrane index, cell caspase-like protease activity, and programmed cell death and hence resulted in increase cell viability. As these isolates remain associated with the roots, the effects of tolerance against salinity are observed here. Results also indicate that isolated PGPR strain help in alleviating up to 1.5 % salinity stress as well as improve tolerance.     

Heat Shock Induces Programmed Cell Death in Wheat Leaves

Programmed cell death (PCD) was triggered in wheat leaves by a heat treatment (42 °C). This treatment caused DNA fragmentation as shown in DNA laddering analysis and in terminal deoxynucleotidyl transferase-mediated deoxyuridine-5-triphosphate (dUTP) nick end labeling (TUNEL) analysis. Methanol and acetone treatment of leaves significantly blocked PCD. Western analysis indicated that a 65 kDa poly(ADP-ribose) polymerase-like protein was degraded during the treatment. However, high temperature (80 °C) treatment caused necrosis but not PCD.     

杜仲次生木质部分化过程中的细胞编程死亡

通过电子显微镜观察、DNA断裂检测及类似半胱氨酸蛋白酶（caspase-like proteases,CLPs)降解检测等技术,对杜仲（Eucommia ulmoides Oliv.）次生木质部分化过程的细胞编程死亡进行了研究。分化中的次生木质部细胞总DNA凝胶电泳检测到DNA ladder,并通过TUNEL检测进一步确定了DNA被降解。Western blot结果表明：caspase-8和caspase-3状蛋白酶（caspase-8-和caspase-3-like proteases,CLPs)及多聚ADP-核糖聚合酶（poly(ADP-ribose) polymerase,PARP)在次生木质部分化过程中被降解。这些研究结果表明,杜仲次生木质部的细胞分化是一个典型的编程性死亡（Programmed cell death,PCD)过程,CLPs可能参与了此过程。