Increasing the Antitumor Effect of an EpCAM-Targeting Fusion Toxin by Facile Click PEGylation

Fusion toxins used for cancer-related therapy have demonstrated short circulation half-lives, which impairs tumor localization and, hence, efficacy. Here, we demonstrate that the pharmacokinetics of a fusion toxin composed of a designed ankyrin repeat protein (DARPin) and domain I–truncated Pseudomonas Exotoxin A (PE40/ETA″) can be significantly improved by facile bioorthogonal conjugation with a polyethylene glycol (PEG) polymer at a unique position. Fusion of the anti-EpCAM DARPin Ec1 to ETA″ and expression in methionine-auxotrophic E. coli enabled introduction of the nonnatural amino acid azidohomoalanine (Aha) at position 1 for strain-promoted click PEGylation. PEGylated Ec1-ETA″ was characterized by detailed biochemical analysis, and its potential for tumor targeting was assessed using carcinoma cell lines of various histotypes in vitro, and subcutaneous and orthotopic tumor xenografts in vivo. The mild click reaction resulted in a well-defined mono-PEGylated product, which could be readily purified to homogeneity. Despite an increased hydrodynamic radius resulting from the polymer, the fusion toxin demonstrated high EpCAM-binding activity and retained cytotoxicity in the femtomolar range. Pharmacologic analysis in mice unveiled an almost 6-fold increase in the elimination half-life (14 vs. 82 minutes) and a more than 7-fold increase in the area under the curve (AUC) compared with non-PEGylated Ec1-ETA″, which directly translated in increased and longer-lasting effects on established tumor xenografts. Our data underline the great potential of combining the inherent advantages of the DARPin format with bioorthogonal click chemistry to overcome the limitations of engineering fusion toxins with enhanced efficacy for cancer-related therapy.

Liquid-Crystalline Dendrimers Designed by Click Chemistry

Liquid-crystalline dendrimers have been prepared from second-generation Percec-type poly(benzyl ether) dendrons or second-generation poly(aryl ester) dendrons carrying cyanobiphenyl mesogens. The Janus dendrimer, which combines the two types of dendromesogens, has also been synthesized. Those compounds have been prepared under copper-catalyzed azide–alkyne cycloaddition conditions. The mesomorphic properties have been studied by thermal analysis (POM, DSC) and small-angle X-ray scattering. Smectic A, nematic, and columnar phases have been observed depending on the dendritic building blocks. The click reaction has proven to be a powerful and elegant synthetic tool for the design of complex dendritic liquid-crystalline architectures.

Real time monitoring of click chemistry self-healing in polymer composites

A photo-healable rubber composite based on effective and fast thiol-alkyne click chemistry as a selfhealing agent prestored in glass capillaries is reported. The click reaction and its effect on the mechanical properties of the composite are monitored in real time by dynamic mechanical analysis, showing that the successful bleeding of healing agents to the crack areas and the effective photoinitiated click reaction result in a 30% storage modulus increase after only 5 min of UV light exposure. X-ray tomography confirms capillary-driven bleeding of reactants to the damaged areas. The effect of storing the click chemistry reactants in separate capillaries is also studied, and results show the importance of stoichiometry in achieving a significant level of repair of the composite. No reactant degradation or premature chemical reaction is observed over time in samples stored in the absence of UV radiation; they are able to undergo the self-healing reaction even one month after preparation.

Have a heart... click from the start!.

Mode of access: Internet.

"Click it or Ticket" program evaluation : June 2002 observational survey results / compiled and prepared by Illinois Dept. of Transportation, Division of Traffic Safety, Evaluation Unit.

"August 2002"--Cover.

Evaluation of the 2006 Illinois "Click It or Ticket" Campaign, April 24-June 18, 2006 /

Evaluates the impact of the "Click It or Ticket" campaign (May 2006) by using statewide public opinion and observational surveys of licensed Illinois drivers.

Evaluation of the 2006 Thanksgiving Click It or Ticket Campaign in Illinois, November 6 - December 10, 2006 /

Evaluates the impact of the "Click It or Ticket" campaign (April 2007) by using statewide public opinion and observational surveys of licensed Illinois drivers.

Motorist awareness and opinion survey on safety belt issues during the "click it or ticket" campaign in Illinois /

"June 2002 survey results"

Cyber-rigging click-through rates: Exploring the ethical dimensions

Click-through rate is considered a very important metric and a key performance indicator of the success of online advertising and is the most frequently used measure to gauge the effectiveness of banner advertising. Marketers also use click-through rates in arriving at performance measurement activities such as the calculation of 'customer life time value' and 'customer acquisition cost'. Click-through is the second most frequently used banner ad pricing method after cost per thousand impressions. Online advertising is facing a new form of challenge – the artificial inflation of click-through rates. We call this practice 'cyber-rigging'. The objective of this paper is to explore the ethical dimensions of cyber-rigging through application of ethical principles and theories.

Poly(3-hexylthiophene) based block copolymers prepared by "click" chemistry

p-Conjugated block copolymers have been prepared from terminal azide functionalized polystyrenes (PS) and alkyne functionalized poly(3- hexylthiophene)s (P3HT) via a copper(I) catalyzed Huisgen [3 + 2] dipolar cycloaddition reaction. The functionalized a-azido-PS homopolymer was prepared by atom transfer radical polymerization from a specifically designed initiator bearing the azide function, whereas ?-ethynyl-P3HT and a,?-pentynyl-P3HT were synthesized by a modified Grignard metathesis polymerization using alkynyl Grignard derivatives. The electronic environment of the alkynyl end groups was shown to be decisive in determining triazole ring formation.

A lifetime of language experience at each click!

Native speakers learn their mother tongue slowly, from birth, by the constant repetition of common words and phrases in a variety of contexts and situations, within the language community. As foreign language learners, we face considerable disadvantages when compared to children learning their mother tongue. Foreign language learners start later in life, have less time, have fewer opportunities to experience the language, and learn in the restricted environment of the classroom. Teachers and books give us information about many words and phrases, but it is difficult for us to know what we need to focus on and learn thoroughly, and what is less important. The rules and explanations are often difficult for us to understand. A large language corpus represents roughly the amount and variety of language that a native-speaker experiences in a whole lifetime. Learners can now access language corpora. We can check which words and phrases are important, and quickly discover their common meanings, collocations, and structural patterns. It is easier to remember things that we find out ourselves, rather than things that teachers or books tell us. Each click on the computer keyboard can show us the same information in different ways, so we can understand it more easily. We can also get many more examples from a corpus. Teachers and native-speakers can also use corpora, to confirm and enhance their own knowledge of a language, and prepare exercises to guide their students. Each of us can learn at our own level and at our own speed.

Facile synthesis of well-defined hydrophilic methacrylic macromonomers using ATRP and click chemistry

A range of well-defined hydrophilic methacrylic macromonomers has been synthesized by the judicious combination of atom transfer radical polymerization (ATRP) and copper-catalyzed 1,3-dipolar cycloaddition (azide-alkyne click chemistry). An azido a-functionalized ATRP initiator was used to produce well-defined homopolymers with terminal azide functionality via ATRP in protic media at 20 °C, with generally good control being achieved over both target molecular weight and final polydispersity (Mw/Mn = 1.10-1.35). Suitable methacrylic monomers include 2-aminoethyl methacrylate hydrochloride, 2-(diethylamino)ethyl methacrylate, 2-(dimethylamino)ethyl methacrylate, 2-hydroxyethyl methacrylate, 2-hydroxypropyl methacrylate, 2-(methacryloyloxy)ethyl phosphorylcholine, glycerol monomethacrylate, potassium 3-sulfopropyl methacrylate, and quaternized 2-(dimethylamino)ethyl methacrylate. These homopolymer precursors were then efficiently clicked using either propargyl methacrylate or propargyl acrylate to yield near-monodisperse (meth)acrylate-capped macromonomers with either cationic, anionic, nonionic, or zwitterionic character. Moreover, this generic route to well-defined hydrophilic macromonomers is also suitable for “one-pot” syntheses, as exemplified for 2-hydroxyethyl methacrylate and glycerol monomethacrylate-based macromonomers.