Iterative in Situ Click Chemistry Assembles a Branched Capture Agent and Allosteric Inhibitor for Akt1

We describe the use of iterative in situ click chemistry to design an Akt-specific branched peptide triligand that is a drop-in replacement for monoclonal antibodies in multiple biochemical assays. Each peptide module in the branched structure makes unique contributions to affinity and/or specificity resulting in a 200 nM affinity ligand that efficiently immunoprecipitates Akt from cancer cell lysates and labels Akt in fixed cells. Our use of a small molecule to preinhibit Akt prior to screening resulted in low micromolar inhibitory potency and an allosteric mode of inhibition, which is evidenced through a series of competitive enzyme kinetic assays. To demonstrate the efficiency and selectivity of the protein-templated in situ click reaction, we developed a novel QPCR-based methodology that enabled a quantitative assessment of its yield. These results point to the potential for iterative in situ click chemistry to generate potent, synthetically accessible antibody replacements with novel inhibitory properties

A New Methodology for Assessing Macromolecular Click Reactions and Its Application to Amine--Tertiary Isocyanate Coupling for Polymer Ligation.

Click reactions have provided access to an array of remarkably complex polymer architectures. However, the term "click" is often applied inaccurately to polymer ligation reactions that fail to respect the criteria that typify a true "click" reaction. With the purpose of providing a universal way to benchmark polymer-polymer coupling efficiency at equimolarity and thus evaluate the fulfilment of click criteria, we report a simple one-pot methodology involving the homodicoupling of α-end-functionalized polymers using a small-molecule bifunctional linker. A combination of SEC analysis and chromatogram deconvolution enables straightforward quantification of the coupling efficiency. We subsequently employ this methodology to evaluate an overlooked candidate for the click reaction family: the addition of primary amines to α-tertiary isocyanates (α-(t)NCO). Using our bifunctional linker coupling strategy, we show that the amine-(t)NCO reaction fulfills the criteria for a polymer-polymer click reaction, achieving rapid, chemoselective, and quantitative coupling at room temperature without generating any byproducts. We demonstrate that amine-(t)NCO coupling is faster and more efficient than the more common amine-tertiary active ester coupling under equivalent conditions. Additionally, we show that the α-(t)NCO end group is unprecedentedly stable in aqueous media. Thus, we propose that the amine-(t)NCO ligation is a powerful new click reaction for efficient macromolecular coupling.Dr Maarten Danial for providing the cyclic peptide.This is the final version of the article. It first appeared from the American Chemical Society via http://dx.doi.org/10.1021/jacs.5b1183

Isolation of bis(copper) key intermediates in Cu-catalyzed azide-alkyne "click reaction".

The copper-catalyzed 1,3-dipolar cycloaddition of an azide to a terminal alkyne (CuAAC) is one of the most popular chemical transformations, with applications ranging from material to life sciences. However, despite many mechanistic studies, direct observation of key components of the catalytic cycle is still missing. Initially, mononuclear species were thought to be the active catalysts, but later on, dinuclear complexes came to the front. We report the isolation of both a previously postulated π,σ-bis(copper) acetylide and a hitherto never-mentioned bis(metallated) triazole complex. We also demonstrate that although mono- and bis-copper complexes promote the CuAAC reaction, the dinuclear species are involved in the kinetically favored pathway

Selective C(sp2)−H Halogenation of "click" 4-Aryl-1,2,3-triazoles

Selective bromination reactions of “click compounds” are described. Electron-neutral and electron-deficient arenes selectively undergo unprecedented Pd-catalyzed C–H ortho-halogenations assisted by simple triazoles as modular directing groups, whereas electron-rich arenes are regioselectively halogenated following an electrophilic aromatic substitution reaction pathway. These C–H halogenation procedures exhibit a wide group tolerance, complement existing bromination procedures, and represent versatile synthetic tools of utmost importance for the late-stage diversification of “click compounds”. The characterization of a triazole-containing palladacycle and density functional theory studies supported the mechanism proposal.We are grateful to Gobierno Vasco (ELKARTEK_KK-2015/0000101; IT_1033-16) and UPV/EHU (GIU15/31) for financial support. A. C. thanks MINECO for a Ramón y Cajal research contract (RYC-2012-09873). Cost-CHAOS action is also acknowledged

Development of mixed metal metal-organic polyhedra networks, colloids, and MOFs and their pharmacokinetic applications

The coordination networking of discrete metal-organic polyhedra (MOPs) involving different ligands as well as metals is a challenging task due to the features of limited solubility and chemical stability of these polyhedra. An unusual approach, ligand-oriented polyhedral networking via click chemistry and further metal coordination is reported here. An alkyne decorated Cu(II)-MOP self-catalyzes the regioselective click reaction (1,3-dipolar cycloaddition) using azide-functionalized ligands under unconventional reaction conditions. Introducing new metal ions, M(II), interlinks the carboxylic groups on the MOP surfaces creating coordination networks. On the other hand, exposure of the respective individual ligand components in the presence of Cu(II) promotes an in-situ click reaction along with metal coordination generating a new 3D-framework. These materials demonstrated a high drug hosting potential exhibiting a controlled progressive release of anticancer (5-flourouracil) and stimulant (caffeine) drugs in physiological saline at 37 degrees C. These innovative and unconventional MOP networks provide a significant conceptual advance in understanding

Templated synthesis of cyclic poly(ionic liquid)s

Charged cyclic polymers, e.g. cyclic DNAs and polypeptides, play enabling roles in organisms, but their synthesis was challenging due to the well known polyelectrolyte effect. To tackle the challenge, we developed a templated method to synthesize a library of imidazolium and pyridinium based cyclic poly(ionic liquid)s. Cyclic templates, cyclic polyimidazole and poly(2-pyridine), were synthesized first through ring-closure method by light-induced Diels-Alder click reaction. Through quaternization of cyclic templates followed by anion metathesis, the cyclic poly(ionic liquid)s were synthesized, which paired with varied counter anions.Comment: 23 pages, 15 figures, 2 table

Triazolinediones as highly enabling synthetic tools

Triazolinediones (TADs) are unique reagents in organic synthesis that have also found wide applications in different research disciplines, in spite of their somewhat "exotic" reputation. In this review, we offer two case studies that demonstrate the possibilities of these versatile and reliable synthetic tools, namely, in the field of polymer science as well as in more recently emerging applications in the field of click chemistry. As the general use of triazolinediones has always been hampered by the limited commercial and synthetic availability of such reagents, we also offer a review of the available TAD reagents, together with a detailed discussion of their synthesis and reactivity. This review thus aims to serve as a practical guide for researchers that are interested in exploiting and further developing the exceptional click -like reactivity of triazolinediones in various applications

Acoustic behavior of melon-headed whales varies on a diel cycle.

Many terrestrial and marine species have a diel activity pattern, and their acoustic signaling follows their current behavioral state. Whistles and echolocation clicks on long-term recordings produced by melon-headed whales (Peponocephala electra) at Palmyra Atoll indicated that these signals were used selectively during different phases of the day, strengthening the idea of nighttime foraging and daytime resting with afternoon socializing for this species. Spectral features of their echolocation clicks changed from day to night, shifting the median center frequency up. Additionally, click received levels increased with increasing ambient noise during both day and night. Ambient noise over a wide frequency band was on average higher at night. The diel adjustment of click features might be a reaction to acoustic masking caused by these nighttime sounds. Similar adaptations have been documented for numerous taxa in response to noise. Or it could be, unrelated, an increase in biosonar source levels and with it a shift in center frequency to enhance detection distances during foraging at night. Call modifications in intensity, directionality, frequency, and duration according to echolocation task are well established for bats. This finding indicates that melon-headed whales have flexibility in their acoustic behavior, and they collectively and repeatedly adapt their signals from day- to nighttime circumstances

Efficient click-addition sequence for polymer–polymer couplings

Controlled radical polymerization methods and click chemistry form a versatile toolbox for creating complex polymer architectures. However, the incompatibility between the functional groups required for click reactions and the reaction conditions of radical polymerization techniques often limits application. Here, we demonstrate how combining two complementary click reactions in a sequence circumvents compatibility issues. We employ isocyanate-amine addition on a polymer obtained by RAFT without purification, thus allowing us to work at exact equimolarity. The addition of commercially available amine-functional azido or strained alkyne compounds, yields orthogonally modified polymers, which can be coupled together in a subsequent strain promoted cycloaddition (SPAAC). The efficiency of this reaction sequence is demonstrated with different acrylate, methacrylate, and acrylamide polymers giving block copolymers in high yield. The resulting diblock copolymers remain active towards RAFT polymerization, thus allowing access to multiblock structures by simple chain extension. The orthogonality of the isocyanate-amine reaction, SPAAC and RAFT polymerization (both in terms of monomer and chain end groups) is a key advantage and offers access to functional and challenging polymer architectures without the need for stringent reaction conditions or laborious intermediate purifications