Development of thrombolytic agents

Despite their widespread use in patients with acute myocardial infarction, all currently available thrombolytic agents suffer from a number of significant limitations, including resistance to reperfusion, the occurrence of acute coronary reocclusion and bleeding complications. Furthermore, the therapeutic use of plasminogen activators as thrombolytic agents requires intravenous infusion of relatively large amounts of material. Therefore, the quest for thrombolytic agents with a higher thrombolytic potency, specific thrombolytic activity and/or a better fibrin-selectivity continues. Several lines of research towards improvement of thrombolytic agents are being explored, including the construction of mutants and variants of plasminogen activators, chimeric plasminogen activators, conjugates of plasminogen activators with monoclonal antibodies, or plasminogen activators from animal or bacterial origin.     

人t-PA溶栓突变体的研究进展

人t-PA在机体循环中的纤溶系统中起重要作用,是一种内源性溶血栓因子,t-PA蛋白分子可直接用于溶栓治疗,但天然的t-PA分子在体内半衰期短,极最被清除,因而限制其广泛应用,根据它的结构特点而改造的一系列t-PA变体分子将成为新一代溶栓药物,在溶栓治疗中广泛应用。     

缺血性脑卒中溶栓药物研究进展

缺血性脑卒中是一种高发病率、高死亡率的重大健康危机。溶栓药物能快速溶解血栓、减少出血副作用、实现血管再通,对缺血性脑卒中治疗起到关键性的作用。重组组织纤溶酶原激活剂(rtPA)是FDA批准的唯一缺血性脑卒中药物,但在临床使用中有诸多限制。近年来,基于tPA的溶栓药物及治疗策略发展迅速,文中结合笔者课题组及目前国内外的相关研究成果,回顾了该领域的最新进展,为新型溶栓药物发展提供科学依据和思路。     

Development of new thrombolytic agents using recombinant DNA technology.

The increasing incidence of thromboembolic diseases has sustained the search for new agents able to stimulate the natural fibrinolytic system. The first generation of antithrombotic agents include bacterial streptokinase and human urine urokinase. Because these molecules lack specificity for the fibrin clot, important efforts have been made to produce, using recombinant DNA technology, agents presenting higher fibrin clot selectivity such as t-PA (tissue-type plasminogen activator) and scu-PA (single chain urokinase-type plasminogen activator). In parallel, several laboratories are presently attempting to create mutants and hybrids plasminogen activators displaying improved thrombolytic properties with respect to the natural molecules. In this paper, we describe briefly the mechanisms of fibrinolysis and the role of the different natural thrombolytic agents. In addition, we review the possibilities of genetic engineering for the production of natural and novel plasminogen activators.     

Exploitation of phage display for the development of anti-cancer agents targeting fibroblast growth factor signaling pathways: New strategies to tackle an old challenge

A tumor is defined as a group of cancer cells and ‘surrounding’ stromal bio-entities. Alongside the extracellular matrix (ECM) in the tumor microenvironment (TME), the stromal cells play key roles in cancer affliction and progression. Carcinoma-associated fibroblasts (CAFs) in the area of the tumor, whether activated or not, dictate the future of tumor cells. The CAFs and corresponding secreted growth factors (GFs), which mediate the crosstalk within the TME, can be targeted in therapies directed at the stroma. The impact of the fibroblast growth factor-fibroblast growth factor receptor (FGF-FGFR) signaling pathway in different kinds of tumors has been explored. Several tyrosine kinase inhibitors (TKIs), monoclonal antibodies (mAbs), and ligand traps targeting the formation of FGF-FGFR complex are in preclinical or early development phases. Moreover, there are numerous studies in the literature reporting the application of phage display technology for the development of peptides and proteins capable of functioning as FGF mimetics or traps, which are able to modulate FGF-related signaling pathways. In this review, prominent research in relation to phage display-assisted ligand identification for the FGF/FGFR system is discussed.     

Current strategies in the search for novel antiparasitic agents

The market for antiparasitic products comprises the largest segment for sales of livestock and companion-animal healthcare agents. Despite the availability of highly effective, broad-spectrum agents, there remains a need for safer, more convenient and more environmentally friendly products that will overcome the ever-present threat of resistance development. The very high cost of discovering and developing a new drug, especially for use in livestock, is reflected in the limited number of new classes of antiparasitic agent launched on the market. New strategies are being adopted to minimise the cost of discovering potential drug candidates by maximising the chance of identifying a useful target mechanism of action and by speeding the time to discover and optimise a lead structure. These rely heavily on new technologies in target identification, screen development and lead optimisation. Examples of these will be discussed and speculation made about the possible factors that could influence the future shape of antiparasitic control.     

Design strategies for anti-amyloid agents

Numerous diseases have been linked to a common pathogenic process called amyloidosis, whereby proteins or peptides clump together in the brain or body to form toxic soluble oligomers and/or insoluble fibres. An attractive strategy to develop therapies for these diseases is therefore to inhibit or reverse protein/peptide aggregation. A diverse range of small organic ligands have been found to act as aggregation inhibitors. Alternatively, the wild-type peptide can be derivatised so that it still binds to the amyloid target, but prevents further aggregation. This can be achieved by adding a bulky group or charged amino acid to either end of the peptide, or by incorporating proline residues or N-methylated amide groups.     

Protamine, a substrate for thrombolytic agents and enzymes of similar specificity.

Protamine, a highly purified basic polypeptide of 4000 molecular weight containing 80–85% arginine, is a useful substrate for the assay of plasmin, activated plasminogen, and enzymes of similar specificity, e.g., urokinase, coagulation factor Xa, trypsin, and thrombin, and is also an excellent secondary substrate for activator assays of urokinase and streptokinase. The assays were performed manually, and also automated procedures for continuous multiple sample analyses were used. The relative sensitivities for various plasmin-like enzymes were: trypsin > plasmin > urokinase > factor Xa > thrombin. Using protamine with manual assay procedures, the amino-terminal groups of the enzyme-degraded protamine digestion products were detected and quantitated by the colorimetric ninhydrin or the fluorometric fluorescamine procedures, and using protamine with an automated system the ninhydrin method was used. Assigning the CTA casein assay for plasmin a nominal sensitivity of 0.1 (for 0.1 CTA unit of plasmin), the sensitivities of the various assay methods were casein, automated protamine, and manual protamine with ninhydrin, 0.1; manual protamine-trichloroacetic acid with fluorescamine, 0.005; and manual protamine direct fluorescamine, 0.0005. A unit of plasmin, based on the uptake of 1 μequiv base/min during hydrolysis of 0.4% protamine sulfate under standard conditions, is equal to approximately 1.7 × 10³ RFI units or 2.9 CTA units; or, 1 CTA unit of plasmin resulted in an average uptake of 0.346 μmol of base or equivalent bonds split per minute.     

Conjugation of plasminogen activators and fibrin-specific antibodies to improve thrombolytic therapeutic agents.

Here we have reviewed chemical and recombinant approaches to the construction of hybrid molecules that combine a "targeting" antibody and an "effector" enzyme activity. There are advantages and disadvantages to both chemical and recombinant methods, and one goal of this review has been to elucidate these so that the appropriate method can be used by those interested in using hybrid molecules to study questions of basic or therapeutic importance. The system studied in greatest detail has as its goal the targeting of a plasminogen activator to an occlusive intravascular thrombus. We have, therefore, used this system as an example of currently available approaches. Now that these methodologies have been studied and put into use, it is anticipated that this principle will be generalized both to other therapeutic applications, as well as to the design and construction of molecules that will allow more basic questions to be addressed.     

低碳经济发展战略研究进展

发展低碳经济已成为国家和区域发展的重要战略。从发展战略的角度,对低碳经济的产生背景、内涵和理论基础等进行了总结和归纳;在此基础上提出了低碳经济发展战略框架研究的主要内容,包括碳政治和碳排放权分配、目标思路、指标体系构建和碳排放的核算;从政策路径、政策工具和政策仿真等方面介绍了低碳经济发展战略的政策研究的主要内容和国内外研究进展。最后指出当前研究的不足之处和未来研究趋向,主要有:低碳经济理论的深入研究,低碳经济战略和政策工具研究,区域低碳经济发展模式研究,低碳经济发展潜力、市场规模和经济技术风险研究,碳减排标准规范的研究和制定。     

Effectiveness of acylated thrombolytic agents in lysis of blood plasma clots from humans and various animals

Kinetics of lysis of fibrin clots from the human, guinea pig, rat and rabbit blood plasma by two active-site-acylated derivatives of the activator plasmin-streptokinase complex with different reaction rate constants has been studied in vitro. It is found that lysis of blood plasma clots in guinea pig is most similar to that of man. Acyl activator dose being increased, the lysis of a plasma clot in guinea pig is accelerated. Two acyl activators exhibit higher fibrinolytic-efficiency as compared to a free activator. Experiments carried out in vivo on guinea pigs with thrombosis show that acyl activators, in contrast to nonmodified plasmin-streptokinase complex induce the less system activation of fibrinolysis and the less fibrinogenolysis.