SET‐LRP of acrylates in the presence of radical inhibitors

The Cu(0)/Me₆‐TREN‐catalyzed single‐electron transfer mediated living radical polymerization (SET‐LRP) of methyl acrylate in the presence of the classic 4‐methoxyphenol free radical inhibitor was investigated. Kinetic experiments, combined with ¹H NMR, and MALDI‐TOF MS analysis of the resulting polyacrylates demonstrated that SET‐LRP is a robust synthetic method that does not require the purification of the monomers to remove the radical inhibitor. It is anticipated that these results will contribute to the expansion of technological and fundamental applications of SET‐LRP since it allows the synthesis of polymers with a structural perfection that previously was not accessible by any other method, starting from unpurified monomers, solvents, initiators, and ligands. © 2008 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 3174–3181, 2008     

Analysis of dimeric cyanine–nucleic acid dyes by capillary zone electrophoresis in N,N-dimethylacetamide as non-aqueous organic solvent

A method based on capillary zone electrophoresis is presented for the determination of the purity of commercial dimeric cyanine dyes (TOTO, YOYO, BOBO, all -1 and -3 species, LOLO-1, POPO-1) that are common as fluorescent probes for nucleic acid staining. These dyes are tetracharged cations, and have a strong tendency to interact with negatively charged centres, where they are rapidly adsorbed, especially from aqueous solutions. Thus anionic sites at the capillary wall must be avoided, and aqueous buffers are not suitable. The method introduced here avoids both complications, using non-aqueous N,N-dimethylacetamide as solvent, and suppressing the dissociation of silanol groups at the capillary surface due to selection of acidic separation conditions (20 mmol/l perchloric acid as background electrolyte). The present method enables the determination of the purity of all 10 dyes in less than 15 min. The selectivity of the method allows separation of at least five main and differentiating a number of unresolved minor contaminants as demonstrated in detail for TOTO-3 as an example. Quantitation (with 100% normalisation of the peak areas) of nine lots of this dye results in a purity between 33 and 87%.     

Synthesis of dendritic macromolecules through divergent iterative thio‐bromo “Click” chemistry and SET‐LRP

The development of a novel nucleophilic thio‐bromo “Click” reaction, specifically base‐mediated thioetherification of thioglycerol with α‐bromoesters was reported in an earlier article. The combination of this thio‐bromo click reaction with subsequent acylation with 2‐bromopropionyl bromide provides an iterative two‐step divergent growth approach to the synthesis of a new class of poly(thioglycerol‐2‐ propionate) (PTP) dendrimers. In this article, the addition of a third step, the single‐electron transfer living radical polymerization (SET‐LRP) of methyl acrylate (MA), was shown to provides access to a three‐step “branch” and “grow” divergent approach to dendritic macromolecules wherein poly(methyl acrylate) (PMA) connects the branching subunits. This facile methodology can provide a diversity of dendritic macromolecular topologies and will ultimately provide the means to the development of self‐organizable dendritic macromolecules. © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47: 3940–3948, 2009     

Alkyl chloride initiators for SET‐LRP of methyl acrylate

Single‐electron transfer living radical polymerization (SET‐LRP) proceeds by an outer‐sphere single‐electron transfer mechanism that induces a heterolytic bond cleavage of the initiating and propagating R‐X (where X = Cl, Br, and I) species. Therefore, unlike the homolytic bond cleavage mechanism claimed for ATRP, SET‐LRP is expected to show a small dependence of the nature of the halide group on the apparent rate constant of activation. This means the R‐X with X = Cl, Br, and I must all be efficient initiators for SET‐LRP and no chain transfer must be observed in the case of initiators with X = Br and I. Here, we report the SET‐LRP of methyl acrylate initiated with the alkyl chlorides methyl‐2‐chloropropionate (MCP) and chloroform (CHCl₃) and catalyzed by Cu(0)/Me₆‐TREN/CuCl₂ in DMSO at 25 °C. A combination of kinetic and structural analysis was used to elucidate the MCP and CHCl₃ initiating behavior under SET‐LRP conditions, and to demonstrate the very small dependence of the SET‐LRP apparent rate constant of propagation on X while providing polymers with well defined architecture. © 2008 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 4917–4926, 2008     

Purification and characterization of recombinant human renin for X-ray crystallization studies

Background  

The renin-angiotensin-aldosterone system (RAS) cascade is a major target for the clinical management of hypertension. Although inhibitors of various components of this cascade have been developed successfully, development of renin inhibitors has proven to be problematic. The development of these inhibitors has been hindered by poor bioavailability and complex synthesis. However, despite the challenges of designing renin inhibitors, the enzyme remains a promising target for the development of novel treatments for hypertension. X-ray crystallographic data could greatly assist the design and development of these inhibitors. Here we describe the purification and characterization of recombinant human renin for x-ray crystallization studies.     

Synthesis and spectroscopic characterization of red-shifted spironaphthoxazine based optical switch probes

Spironaphthoxazine (NISO) is an efficient optical switch probe that has applications in high contrast detection of Förster resonance energy transfer (FRET) using optical lock-in detection (OLID). NISO exists in two distinct states, spiro (SP) and merocyanine (MC), that can be independently controlled by using alternate irradiation with near ultraviolet and visible light. Unfortunately, the SP-state of NISO has an absorption centered at 350 nm, which may lead to phototoxic effects when manipulating the probe within a living cell. To overcome this problem we introduce new, red-shifted amino-substituted NISO probes compared to NISO that undergo an efficient SP to MC transition in response to irradiation by using 405-nm light, which is less damaging to living cells. This study details the synthesis of amino-substituted NISO and their N-hydroxysuccinimide ester and maleimide derivatives and their use in generating covalent attached protein conjugates. This study also presents a characterization of the spectroscopic and optical switching properties of these red-shifted NISO probe in solution.