Synthesis and antibacterial activity of 9-oxime ether non-ketolides,and novel binding mode of alkylides with bacterial rRNA

We report a series of new 9-oxime ether non-ketolides, including 3-hydroxyl, 3-O-acyl and 3-O-alkyl clarithromycin derivatives, and thiophene-containing ketolides 1b–1d. Unlike previously reported ketolide 1a, none of them is comparable to telithromycin. A molecular modeling study was performed to gain insight into the binding mode of alkylides 17–20 with bacterial rRNA and to rationalize the great disparity of their SAR. The 3-O-sidechains of 19 and 20 point to the so-called hydrophilic side of the macrolide ring, as seen in clarithromycin. In contrast, the 3-O-sidechains of 17 and 18 bend to the backside, the so-called hydrophobic side of the macrolide ring. The results clearly indicated the alkylides with improved antibacterial activity might possess a novel binding mode, which is different from clarithromycin and the alkylides with poor activity.     

Synthesis and acrosin inhibitory activities of 5-phenyl-1H-pyrazole-3-carboxylic acid amide derivatives

A series of novel 5-phenyl-1H-pyrazole-3-carboxylic acid amide derivatives were designed, synthesized, and their acrosin inhibitory activities in vitro were evaluated. The results of the acrosin inhibitory activity showed that all target compounds were more potent than control TLCK. Compounds AQ-A1, AQ-D3, AQ-D4, AQ-E4 and AQ-E5 exhibited stronger acrosin inhibitory activities than control ISO-1. Especially, compound AQ-E5 displayed the most potent acrosin inhibitory activity in all the compounds, with an IC₅₀ of 0.01 μmol/mL. This study provided a new structural class for the development of novel acrosin inhibitory agents.     

Genetic diversity of Bartonella quintana in macaques suggests zoonotic origin of trench fever

Bartonella quintana is a bacterium that causes a broad spectrum of diseases in humans including trench fever. Humans were previously considered to be the primary, if not the only, reservoir hosts for B. quintana. To identify the animal reservoir and extend our understanding of the ecological and evolutionary history of B. quintana, we examined blood samples from macaques and performed multilocus sequence typing (MLST) analysis. We demonstrated the prevalence of B. quintana infection was common in macaques from main primate centres in mainland China. Overall, 18.0% (59/328) of rhesus macaques and 12.7% (39/308) of cynomolgus macaques were found to be infected with B. quintana by blood culture and/or polymerase chain reaction. The infection was more frequently identified in juvenile and young monkeys compared with adult animals. In contrast with the relatively low level of sequence divergence of B. quintana reported in humans, our investigation revealed much higher genetic diversity in nonhuman primates. We identified 44 new nucleotide variable sites and 14 novel sequence types (STs) among the B. quintana isolates by MLST analysis. Some STs were found only in cynomolgus macaques, while some others were detected only in rhesus macaques, suggesting evidence of host‐cospeciation, which were further confirmed by phylogenetic analysis and Splits decomposition analysis. Our findings suggest that trench fever may primarily be a zoonotic disease with macaques as the natural hosts.     

Discovery of 3-phenyl-1H-5-pyrazolylamine derivatives containing a urea pharmacophore as potent and efficacious inhibitors of FMS-like tyrosine kinase-3 (FLT3)

Preclinical investigations and early clinical trials suggest that FLT3 inhibitors are a viable therapy for acute myeloid leukemia. However, early clinical data have been underwhelming due to incomplete inhibition of FLT3. We have developed 3-phenyl-1H-5-pyrazolylamine as an efficient template for kinase inhibitors. Structure–activity relationships led to the discovery of sulfonamide, carbamate and urea series of FLT3 inhibitors. Previous studies showed that the sulfonamide 4 and carbamate 5 series were potent and selective FLT3 inhibitors with good in vivo efficacy. Herein, we describe the urea series, which we found to be potent inhibitors of FLT3 and VEGFR2. Some inhibited growth of FLT3-mutated MOLM-13 cells more strongly than the FLT3 inhibitors sorafenib (2) and ABT-869 (3). In preliminary in vivo toxicity studies of the four most active compounds, 10f was found to be the least toxic. A further in vivo efficacy study demonstrated that 10f achieved complete tumor regression in a higher proportion of MOLM-13 xenograft mice than 4 and 5 (70% vs 10% and 40%). These results show that compound 10f possesses improved pharmacologic and selectivity profiles and could be more effective than previously disclosed FLT3 inhibitors in the treatment of acute myeloid leukemia.     

Functional consequences of retro-inverso isomerization of a miniature protein inhibitor of the p53–MDM2 interaction

Peptide retro-inverso isomerization is thought to be functionally neutral and has been widely used as a tool for designing proteolytically stable d-isomers to recapitulate biological activities of their parent l-peptides. Despite success in a wide range of applications, exceptions amply exist that clearly defy this rule of thumb when parent l-peptides adopt an α-helical conformation in their bound state. The detrimental energetic effect of retro-inverso isomerization of an α-helical l-peptide on its target protein binding has been estimated to be 3.0–3.4 kcal/mol. To better understand how the retro-inverso isomer of a structured protein works at the molecular level, we chemically synthesized and functionally characterized the retro-inverso isomer of a rationally designed miniature protein termed stingin of 18 amino acid residues, which adopts an N-terminal loop and a C-terminal α-helix stabilized by two intra-molecular disulfide bridges. Stingin emulated the transactivation peptide of the p53 tumor suppressor protein and bound with high affinity and via its C-terminal α-helix to MDM2 and MDMX—the two negative regulators of p53. We also prepared the retro isomer and d-enantiomer of stingin for comparative functional studies using fluorescence polarization and surface plasmon resonance techniques. We found that retro-inverso isomerization of l-stingin weakened its MDM2 binding by 720 fold (3.9 kcal/mol); while enantiomerization of l-stingin drastically reduced its binding to MDM2 by three orders of magnitude, sequence reversal completely abolished it. Our findings demonstrate the limitation of peptide retro-inverso isomerization in molecular mimicry and reinforce the notion that the strategy works poorly with biologically active α-helical peptides due to inherent differences at the secondary and tertiary structural levels between an l-peptide and its retro-inverso isomer despite their similar side chain topologies at the primary structural level.¹     

The discovery of colchicine-SAHA hybrids as a new class of antitumor agents

A novel class of colchicine-SAHA hybrids were designed and synthesised based on the synergistic antitumor effect of tubulin inhibitors and histone deacetylases (HDAC) inhibitors. To the best of our knowledge, this is the first design of molecules that are dual inhibitors of tubulin and HDAC. Biological evaluations of these compounds included the inhibitory activity of HDAC, in vitro cell cycle analysis in BEL-7402 cells as well as cytotoxicity in five cancer cell lines.     

Hexamers of the Type II Secretion ATPase GspE from Vibrio cholerae with Increased ATPase Activity

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艾比湖地区土壤呼吸对季节性冻土厚度变化的响应

利用LI-8100土壤CO2排放通量全自动测量系统,于2010年1～4月测定了艾比湖地区不同植被类型样地的土壤呼吸速率,结合环境因子、冻土厚度及室内土壤理化性质分析,探讨了温带干旱区季节性冻土厚度变化对土壤呼吸的影响。结果表明：土壤温度在冻结期是影响冻土厚度的最主要环境因子,而解冻期冻土厚度变化与土壤温度等环境因子关系不显著（P>0.05）;冻土厚度在不同时期影响土壤呼吸速率的程度不同,冻结期两者呈显著正相关（R2=0.782,P<0.05）,解冻初期两者呈弱相关（P>0.05）;土壤呼吸速率在土壤冻结期与解冻初期不存在显著差异（P>0.05）,但在解冻完全期则表现出明显的增加趋势（差值为0.14～0.37μmol?m-2?s-1）,表明冻土融化会明显地增加土壤碳排放,从而增加大气中的CO2。研究结果初步阐明了艾比湖地区季节性冻土厚度变化对土壤呼吸的影响,为揭示全球变暖背景下冻土退化过程中的碳释放机理提供理论基础。     

基于盐分梯度的荒漠植物多样性与群落、种间联接响应

土壤盐分是影响干旱区荒漠植物群落动态的决定因素之一。基于样方调查和不同土壤盐分梯度下植物多样性指数及群落与种间关联的计算结果,分析干旱区荒漠群落植物多样性、群落联接性和种间关联对土壤盐分梯度的响应动态。结果表明,在土壤盐含量为0.03%-0.55% (S1)、0.61%-1.24% (S2)和1.41%-1.79% (S3)的盐分梯度上,(1)随土壤盐含量升高,群落生活型结构改变,草本比例减少,乔木比例增加;(2)植物多样性指数随土壤盐分增加而下降,低盐分梯度下,二者极显著正相关(P<0.01),中盐梯度下二者间呈部分显著负相关(P<0.05),高盐梯度下则转为以正相关为主(P>0.05);(3)群落联接性随土壤盐分梯度转变,在0.61%-1.24%的中度盐含量时正联接性最强(VR=1.89),高盐度含量下群落转为不显著的负联接,稳定性降低;(4)沿盐分升高梯度,种间的负相关种对数增加,正相关种对数减少,种间关联(相关)强度提高,正负相关种对比(PNR)与多样性指数呈显著正相关(P<0.05)。综上可知,干旱区荒漠植物多样性在土壤盐含量达到1.41%-1.79%水平时总体显著降低;土壤盐分水平显著影响植物群落和种间联接性,种间互利性随盐分梯度增加下降,物种趋于独立分布,并最终导致荒漠植物多样性降低。